%PDF-1.3
1 0 obj
<< /Type /Catalog
/Outlines 2 0 R
/Pages 3 0 R >>
endobj
2 0 obj
<< /Type /Outlines /Count 0 >>
endobj
3 0 obj
<< /Type /Pages
/Kids [6 0 R
146 0 R
198 0 R
232 0 R
276 0 R
278 0 R
]
/Count 6
/Resources <<
/ProcSet 4 0 R
/Font << 
/F1 8 0 R
/F2 9 0 R
/F3 10 0 R
/F4 11 0 R
/F5 148 0 R
>>
/XObject << 
/I1 12 0 R
/I2 13 0 R
/I3 165 0 R
/I4 208 0 R
/I5 211 0 R
>>
>>
/MediaBox [0.000 0.000 612.000 792.000]
 >>
endobj
4 0 obj
[/PDF /Text /ImageC ]
endobj
5 0 obj
<<
/Creator (DOMPDF)
/CreationDate (D:20180723000355+00'00')
/ModDate (D:20180723000355+00'00')
/Title (Human satellite cells:  identification on human muscle fibres � PLOS Currents Muscular Dystrophy)
>>
endobj
6 0 obj
<< /Type /Page
/Parent 3 0 R
/Annots [ 14 0 R 16 0 R 18 0 R 20 0 R 22 0 R 24 0 R 26 0 R 28 0 R 30 0 R 32 0 R 34 0 R 36 0 R 38 0 R 40 0 R 42 0 R 44 0 R 46 0 R 48 0 R 50 0 R 52 0 R 54 0 R 56 0 R 58 0 R 60 0 R 62 0 R 64 0 R 66 0 R 68 0 R 70 0 R 72 0 R 74 0 R 76 0 R 78 0 R 80 0 R 82 0 R 84 0 R 86 0 R 88 0 R 90 0 R 92 0 R 94 0 R 96 0 R 98 0 R 100 0 R 102 0 R 104 0 R 106 0 R 108 0 R 110 0 R 112 0 R 114 0 R 116 0 R 118 0 R 120 0 R 122 0 R 124 0 R 126 0 R 128 0 R 130 0 R 132 0 R 134 0 R 136 0 R 138 0 R 140 0 R 142 0 R 144 0 R ]
/Contents 7 0 R
>>
endobj
7 0 obj
<<
/Length 27137 >>
stream

q
375.000 0 0 39.000 222.000 738.000 cm /I2 Do
Q
q
15.000 709.302 577.500 28.698 re W n
0.267 0.267 0.267 rg
BT 15.000 718.042 Td /F2 21.0 Tf  [(Human satellite cells: identification on human muscle fibres)] TJ ET
Q
0.271 0.267 0.267 rg
BT 15.000 700.036 Td /F3 9.8 Tf  [(January 19, 2012)] TJ ET
BT 26.250 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
BT 31.671 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 37.092 688.195 Td /F1 9.8 Tf  [(Luisa Boldrin)] TJ ET
0.271 0.267 0.267 rg
BT 93.447 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 98.868 688.195 Td /F1 9.8 Tf  [(Jennifer E Morgan)] TJ ET
0.271 0.267 0.267 rg
BT 26.250 676.290 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE. Human satellite cells: identification on human muscle fibres. PLOS Currents Muscular Dystrophy. 2012 )] TJ ET
BT 26.250 664.386 Td /F1 9.8 Tf  [(Jan 19 . Edition 1. doi: 10.1371/currents.RRN1294.)] TJ ET
q
15.000 20.421 577.500 641.584 re W n
0.271 0.267 0.267 rg
BT 26.250 635.283 Td /F4 12.0 Tf  [(Abstract)] TJ ET
BT 26.250 615.329 Td /F1 9.8 Tf  [(Satellite cells, normally quiescent underneath the myofibre basal lamina, are skeletal muscle stem cells responsible for )] TJ ET
BT 26.250 603.424 Td /F1 9.8 Tf  [(postnatal muscle growth, repair and regeneration. Since their scarcity and small size have limited study on transverse muscle )] TJ ET
BT 26.250 591.519 Td /F1 9.8 Tf  [(sections, techniques to isolate individual myofibres, bearing their attendant satellite cells, were developed. Studies on mouse )] TJ ET
BT 26.250 579.615 Td /F1 9.8 Tf  [(myofibres have generated much information on satellite cells, but the limited availability and small size of human muscle )] TJ ET
BT 26.250 567.710 Td /F1 9.8 Tf  [(biopsies have hampered equivalent studies of satellite cells on human myofibres. Here, we identified satellite cells on fragments )] TJ ET
BT 26.250 555.805 Td /F1 9.8 Tf  [(of human and mouse myofibres, using a method applicable to small muscle biopsies.)] TJ ET
BT 26.250 519.203 Td /F4 12.0 Tf  [(Funding Statement)] TJ ET
BT 26.250 499.248 Td /F1 9.8 Tf  [(LB is founded by the Muscular Dystrophy Campaign \(MDC\) and JM holds a Wellcome Trust University award.)] TJ ET
BT 26.250 470.146 Td /F4 12.0 Tf  [(Introduction)] TJ ET
BT 26.250 450.192 Td /F1 9.8 Tf  [(Satellite cells are the principal stem cells of skeletal muscle, responsible for its homeostasis, maintenance, growth and )] TJ ET
BT 26.250 438.287 Td /F1 9.8 Tf  [(regeneration through adulthood)] TJ ET
0.267 0.267 0.267 rg
BT 162.292 438.287 Td /F1 9.8 Tf  [([1])] TJ ET
0.271 0.267 0.267 rg
BT 173.134 438.287 Td /F1 9.8 Tf  [( . Satellite cells are located in a niche between the basal lamina and sarcolemma of muscle )] TJ ET
BT 26.250 426.382 Td /F1 9.8 Tf  [(fibres )] TJ ET
0.267 0.267 0.267 rg
BT 52.799 426.382 Td /F1 9.8 Tf  [([2])] TJ ET
0.271 0.267 0.267 rg
BT 63.641 426.382 Td /F1 9.8 Tf  [( . Their rarity and small size are limitations for studying them on transverse muscle sections; therefore techniques were )] TJ ET
BT 26.250 414.477 Td /F1 9.8 Tf  [(developed to prepare isolated muscle fibres, bearing their attendant satellite cells, from rodent muscles)] TJ ET
0.267 0.267 0.267 rg
BT 469.524 414.477 Td /F1 9.8 Tf  [( [3] )] TJ ET
BT 485.787 414.477 Td /F1 9.8 Tf  [([4] )] TJ ET
0.271 0.267 0.267 rg
BT 499.339 414.477 Td /F1 9.8 Tf  [(. These isolated, )] TJ ET
BT 26.250 402.573 Td /F1 9.8 Tf  [(viable muscle fibres, particularly when prepared from genetically-modified mice in which either myonuclei, or satellite cells, )] TJ ET
BT 26.250 390.668 Td /F1 9.8 Tf  [(express a reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 390.668 Td /F1 9.8 Tf  [([5] [6] [7] ,)] TJ ET
0.271 0.267 0.267 rg
BT 175.834 390.668 Td /F1 9.8 Tf  [( have provided a model that has facilitated discovery of novel satellite cell markers, factors )] TJ ET
BT 26.250 378.763 Td /F1 9.8 Tf  [(controlling satellite cell quiescence, activation, proliferation, differentiation, self-renewal and determination of their in vivo )] TJ ET
BT 26.250 366.858 Td /F1 9.8 Tf  [(regenerative capacity )] TJ ET
0.267 0.267 0.267 rg
BT 121.624 366.858 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 132.466 366.858 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 347.454 Td /F1 9.8 Tf  [(Quiescent satellite cells on isolated mouse muscle fibres express Pax7 )] TJ ET
0.267 0.267 0.267 rg
BT 334.574 347.454 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 345.416 347.454 Td /F1 9.8 Tf  [( , M-cadherin )] TJ ET
0.267 0.267 0.267 rg
BT 405.018 347.454 Td /F1 9.8 Tf  [([10])] TJ ET
0.271 0.267 0.267 rg
BT 421.281 347.454 Td /F1 9.8 Tf  [( , CD34, Myf5 )] TJ ET
0.267 0.267 0.267 rg
BT 483.593 347.454 Td /F1 9.8 Tf  [([11] )] TJ ET
0.271 0.267 0.267 rg
BT 502.567 347.454 Td /F1 9.8 Tf  [(caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 549.708 347.454 Td /F1 9.8 Tf  [([12] )] TJ ET
BT 26.250 335.549 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 42.513 335.549 Td /F1 9.8 Tf  [( , calcitonin receptor )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 335.549 Td /F1 9.8 Tf  [([13] [14] )] TJ ET
0.271 0.267 0.267 rg
BT 170.413 335.549 Td /F1 9.8 Tf  [(, nestin )] TJ ET
0.267 0.267 0.267 rg
BT 204.558 335.549 Td /F1 9.8 Tf  [([15] )] TJ ET
0.271 0.267 0.267 rg
BT 223.531 335.549 Td /F1 9.8 Tf  [(and ?7-integrin )] TJ ET
0.267 0.267 0.267 rg
BT 291.274 335.549 Td /F1 9.8 Tf  [([13] [16])] TJ ET
0.271 0.267 0.267 rg
BT 326.511 335.549 Td /F1 9.8 Tf  [( . During activation, myogenic regulatory factors, such as )] TJ ET
BT 26.250 323.644 Td /F1 9.8 Tf  [(MyoD and myogenin, are expressed, and satellite cells either self-renew or differentiate )] TJ ET
0.267 0.267 0.267 rg
BT 405.028 323.644 Td /F1 9.8 Tf  [([8] [17] [18] [19] [20])] TJ ET
0.271 0.267 0.267 rg
BT 491.764 323.644 Td /F1 9.8 Tf  [( . However, both in )] TJ ET
BT 26.250 311.739 Td /F1 9.8 Tf  [(vivo and in vitro studies have demonstrated that the population of mouse satellite cells is heterogeneous, some being more )] TJ ET
BT 26.250 299.835 Td /F1 9.8 Tf  [(�stem cell� like \(i.e. able to regenerate skeletal muscle and functionally reconstitute the satellite cell compartment\) than others )] TJ ET
0.267 0.267 0.267 rg
BT 26.250 287.930 Td /F1 9.8 Tf  [([21] [22] [23] [24])] TJ ET
0.271 0.267 0.267 rg
BT 99.433 287.930 Td /F1 9.8 Tf  [( \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 157.407 287.930 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 168.249 287.930 Td /F1 9.8 Tf  [(\).)] TJ ET
BT 26.250 268.525 Td /F1 9.8 Tf  [(Unfortunately, due to the difficulty in obtaining human muscle in a size and state adequate to allow the preparation of viable )] TJ ET
BT 26.250 256.620 Td /F1 9.8 Tf  [(fibres, there is a gap in our knowledge of human, compared to mouse, satellite cells \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 444.593 256.620 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 455.435 256.620 Td /F1 9.8 Tf  [( \). As a prelude to studies )] TJ ET
BT 26.250 244.716 Td /F1 9.8 Tf  [(on quiescent human satellite cells, we wished to determine antibodies that can be reliably used for their identification. To do )] TJ ET
BT 26.250 232.811 Td /F1 9.8 Tf  [(this, we exploited a technique to isolate fragments of human fibres from muscle biopsies of a normal individual and a Duchenne )] TJ ET
BT 26.250 220.906 Td /F1 9.8 Tf  [(muscular dystrophy \(DMD\) patient. As controls, myofibre fragments from mouse muscles were prepared by the same method. )] TJ ET
BT 26.250 209.001 Td /F1 9.8 Tf  [(We show that Pax7 and M-cadherin are reliable markers of not only mouse, but also human, satellite cells. In contrast, human )] TJ ET
BT 26.250 197.097 Td /F1 9.8 Tf  [(satellite cells are not stained by an antibody to CD34 that recognizes mouse satellite cells and only a subpopulation of human, )] TJ ET
BT 26.250 185.192 Td /F1 9.8 Tf  [(whereas all mouse, satellite cells express caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 254.907 185.192 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 271.170 185.192 Td /F1 9.8 Tf  [(. Conversely, an antibody specific to human ?1 integrin \(CD29\) )] TJ ET
BT 26.250 173.287 Td /F1 9.8 Tf  [(identifies human, but not mouse, satellite cells. We therefore conclude that Pax7 marker defines the majority of not only mouse, )] TJ ET
BT 26.250 161.382 Td /F1 9.8 Tf  [(but also human, satellite cells.)] TJ ET
BT 26.250 124.780 Td /F4 12.0 Tf  [(Materials and Methods)] TJ ET
BT 26.250 104.826 Td /F4 9.8 Tf  [(Ethics)] TJ ET
BT 26.250 85.421 Td /F1 9.8 Tf  [(Tissue sampling was approved by the Institutional Ethics Committee in compliance with national guidelines regarding the use of )] TJ ET
BT 26.250 73.516 Td /F1 9.8 Tf  [(biopsy tissue for research. Patient tissues were collected after written informed consent. Mice were bred and experimental )] TJ ET
BT 26.250 61.611 Td /F1 9.8 Tf  [(procedures were carried out in the Biological Services Unit, University College London, in accordance with the Animals )] TJ ET
BT 26.250 49.707 Td /F1 9.8 Tf  [(\(Scientific Procedures\) Act 1986.)] TJ ET
Q
q
15.000 709.302 577.500 28.698 re W n
0.267 0.267 0.267 rg
BT 15.000 718.042 Td /F2 21.0 Tf  [(Human satellite cells: identification on human muscle fibres)] TJ ET
Q
0.271 0.267 0.267 rg
BT 15.000 700.036 Td /F3 9.8 Tf  [(January 19, 2012)] TJ ET
BT 26.250 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
BT 31.671 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 37.092 688.195 Td /F1 9.8 Tf  [(Luisa Boldrin)] TJ ET
0.271 0.267 0.267 rg
BT 93.447 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 98.868 688.195 Td /F1 9.8 Tf  [(Jennifer E Morgan)] TJ ET
0.271 0.267 0.267 rg
BT 26.250 676.290 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE. Human satellite cells: identification on human muscle fibres. PLOS Currents Muscular Dystrophy. 2012 )] TJ ET
BT 26.250 664.386 Td /F1 9.8 Tf  [(Jan 19 . Edition 1. doi: 10.1371/currents.RRN1294.)] TJ ET
q
15.000 20.421 577.500 641.584 re W n
0.271 0.267 0.267 rg
BT 26.250 635.283 Td /F4 12.0 Tf  [(Abstract)] TJ ET
BT 26.250 615.329 Td /F1 9.8 Tf  [(Satellite cells, normally quiescent underneath the myofibre basal lamina, are skeletal muscle stem cells responsible for )] TJ ET
BT 26.250 603.424 Td /F1 9.8 Tf  [(postnatal muscle growth, repair and regeneration. Since their scarcity and small size have limited study on transverse muscle )] TJ ET
BT 26.250 591.519 Td /F1 9.8 Tf  [(sections, techniques to isolate individual myofibres, bearing their attendant satellite cells, were developed. Studies on mouse )] TJ ET
BT 26.250 579.615 Td /F1 9.8 Tf  [(myofibres have generated much information on satellite cells, but the limited availability and small size of human muscle )] TJ ET
BT 26.250 567.710 Td /F1 9.8 Tf  [(biopsies have hampered equivalent studies of satellite cells on human myofibres. Here, we identified satellite cells on fragments )] TJ ET
BT 26.250 555.805 Td /F1 9.8 Tf  [(of human and mouse myofibres, using a method applicable to small muscle biopsies.)] TJ ET
BT 26.250 519.203 Td /F4 12.0 Tf  [(Funding Statement)] TJ ET
BT 26.250 499.248 Td /F1 9.8 Tf  [(LB is founded by the Muscular Dystrophy Campaign \(MDC\) and JM holds a Wellcome Trust University award.)] TJ ET
BT 26.250 470.146 Td /F4 12.0 Tf  [(Introduction)] TJ ET
BT 26.250 450.192 Td /F1 9.8 Tf  [(Satellite cells are the principal stem cells of skeletal muscle, responsible for its homeostasis, maintenance, growth and )] TJ ET
BT 26.250 438.287 Td /F1 9.8 Tf  [(regeneration through adulthood)] TJ ET
0.267 0.267 0.267 rg
BT 162.292 438.287 Td /F1 9.8 Tf  [([1])] TJ ET
0.271 0.267 0.267 rg
BT 173.134 438.287 Td /F1 9.8 Tf  [( . Satellite cells are located in a niche between the basal lamina and sarcolemma of muscle )] TJ ET
BT 26.250 426.382 Td /F1 9.8 Tf  [(fibres )] TJ ET
0.267 0.267 0.267 rg
BT 52.799 426.382 Td /F1 9.8 Tf  [([2])] TJ ET
0.271 0.267 0.267 rg
BT 63.641 426.382 Td /F1 9.8 Tf  [( . Their rarity and small size are limitations for studying them on transverse muscle sections; therefore techniques were )] TJ ET
BT 26.250 414.477 Td /F1 9.8 Tf  [(developed to prepare isolated muscle fibres, bearing their attendant satellite cells, from rodent muscles)] TJ ET
0.267 0.267 0.267 rg
BT 469.524 414.477 Td /F1 9.8 Tf  [( [3] )] TJ ET
BT 485.787 414.477 Td /F1 9.8 Tf  [([4] )] TJ ET
0.271 0.267 0.267 rg
BT 499.339 414.477 Td /F1 9.8 Tf  [(. These isolated, )] TJ ET
BT 26.250 402.573 Td /F1 9.8 Tf  [(viable muscle fibres, particularly when prepared from genetically-modified mice in which either myonuclei, or satellite cells, )] TJ ET
BT 26.250 390.668 Td /F1 9.8 Tf  [(express a reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 390.668 Td /F1 9.8 Tf  [([5] [6] [7] ,)] TJ ET
0.271 0.267 0.267 rg
BT 175.834 390.668 Td /F1 9.8 Tf  [( have provided a model that has facilitated discovery of novel satellite cell markers, factors )] TJ ET
BT 26.250 378.763 Td /F1 9.8 Tf  [(controlling satellite cell quiescence, activation, proliferation, differentiation, self-renewal and determination of their in vivo )] TJ ET
BT 26.250 366.858 Td /F1 9.8 Tf  [(regenerative capacity )] TJ ET
0.267 0.267 0.267 rg
BT 121.624 366.858 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 132.466 366.858 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 347.454 Td /F1 9.8 Tf  [(Quiescent satellite cells on isolated mouse muscle fibres express Pax7 )] TJ ET
0.267 0.267 0.267 rg
BT 334.574 347.454 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 345.416 347.454 Td /F1 9.8 Tf  [( , M-cadherin )] TJ ET
0.267 0.267 0.267 rg
BT 405.018 347.454 Td /F1 9.8 Tf  [([10])] TJ ET
0.271 0.267 0.267 rg
BT 421.281 347.454 Td /F1 9.8 Tf  [( , CD34, Myf5 )] TJ ET
0.267 0.267 0.267 rg
BT 483.593 347.454 Td /F1 9.8 Tf  [([11] )] TJ ET
0.271 0.267 0.267 rg
BT 502.567 347.454 Td /F1 9.8 Tf  [(caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 549.708 347.454 Td /F1 9.8 Tf  [([12] )] TJ ET
BT 26.250 335.549 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 42.513 335.549 Td /F1 9.8 Tf  [( , calcitonin receptor )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 335.549 Td /F1 9.8 Tf  [([13] [14] )] TJ ET
0.271 0.267 0.267 rg
BT 170.413 335.549 Td /F1 9.8 Tf  [(, nestin )] TJ ET
0.267 0.267 0.267 rg
BT 204.558 335.549 Td /F1 9.8 Tf  [([15] )] TJ ET
0.271 0.267 0.267 rg
BT 223.531 335.549 Td /F1 9.8 Tf  [(and ?7-integrin )] TJ ET
0.267 0.267 0.267 rg
BT 291.274 335.549 Td /F1 9.8 Tf  [([13] [16])] TJ ET
0.271 0.267 0.267 rg
BT 326.511 335.549 Td /F1 9.8 Tf  [( . During activation, myogenic regulatory factors, such as )] TJ ET
BT 26.250 323.644 Td /F1 9.8 Tf  [(MyoD and myogenin, are expressed, and satellite cells either self-renew or differentiate )] TJ ET
0.267 0.267 0.267 rg
BT 405.028 323.644 Td /F1 9.8 Tf  [([8] [17] [18] [19] [20])] TJ ET
0.271 0.267 0.267 rg
BT 491.764 323.644 Td /F1 9.8 Tf  [( . However, both in )] TJ ET
BT 26.250 311.739 Td /F1 9.8 Tf  [(vivo and in vitro studies have demonstrated that the population of mouse satellite cells is heterogeneous, some being more )] TJ ET
BT 26.250 299.835 Td /F1 9.8 Tf  [(�stem cell� like \(i.e. able to regenerate skeletal muscle and functionally reconstitute the satellite cell compartment\) than others )] TJ ET
0.267 0.267 0.267 rg
BT 26.250 287.930 Td /F1 9.8 Tf  [([21] [22] [23] [24])] TJ ET
0.271 0.267 0.267 rg
BT 99.433 287.930 Td /F1 9.8 Tf  [( \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 157.407 287.930 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 168.249 287.930 Td /F1 9.8 Tf  [(\).)] TJ ET
BT 26.250 268.525 Td /F1 9.8 Tf  [(Unfortunately, due to the difficulty in obtaining human muscle in a size and state adequate to allow the preparation of viable )] TJ ET
BT 26.250 256.620 Td /F1 9.8 Tf  [(fibres, there is a gap in our knowledge of human, compared to mouse, satellite cells \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 444.593 256.620 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 455.435 256.620 Td /F1 9.8 Tf  [( \). As a prelude to studies )] TJ ET
BT 26.250 244.716 Td /F1 9.8 Tf  [(on quiescent human satellite cells, we wished to determine antibodies that can be reliably used for their identification. To do )] TJ ET
BT 26.250 232.811 Td /F1 9.8 Tf  [(this, we exploited a technique to isolate fragments of human fibres from muscle biopsies of a normal individual and a Duchenne )] TJ ET
BT 26.250 220.906 Td /F1 9.8 Tf  [(muscular dystrophy \(DMD\) patient. As controls, myofibre fragments from mouse muscles were prepared by the same method. )] TJ ET
BT 26.250 209.001 Td /F1 9.8 Tf  [(We show that Pax7 and M-cadherin are reliable markers of not only mouse, but also human, satellite cells. In contrast, human )] TJ ET
BT 26.250 197.097 Td /F1 9.8 Tf  [(satellite cells are not stained by an antibody to CD34 that recognizes mouse satellite cells and only a subpopulation of human, )] TJ ET
BT 26.250 185.192 Td /F1 9.8 Tf  [(whereas all mouse, satellite cells express caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 254.907 185.192 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 271.170 185.192 Td /F1 9.8 Tf  [(. Conversely, an antibody specific to human ?1 integrin \(CD29\) )] TJ ET
BT 26.250 173.287 Td /F1 9.8 Tf  [(identifies human, but not mouse, satellite cells. We therefore conclude that Pax7 marker defines the majority of not only mouse, )] TJ ET
BT 26.250 161.382 Td /F1 9.8 Tf  [(but also human, satellite cells.)] TJ ET
BT 26.250 124.780 Td /F4 12.0 Tf  [(Materials and Methods)] TJ ET
BT 26.250 104.826 Td /F4 9.8 Tf  [(Ethics)] TJ ET
BT 26.250 85.421 Td /F1 9.8 Tf  [(Tissue sampling was approved by the Institutional Ethics Committee in compliance with national guidelines regarding the use of )] TJ ET
BT 26.250 73.516 Td /F1 9.8 Tf  [(biopsy tissue for research. Patient tissues were collected after written informed consent. Mice were bred and experimental )] TJ ET
BT 26.250 61.611 Td /F1 9.8 Tf  [(procedures were carried out in the Biological Services Unit, University College London, in accordance with the Animals )] TJ ET
BT 26.250 49.707 Td /F1 9.8 Tf  [(\(Scientific Procedures\) Act 1986.)] TJ ET
Q
q
15.000 709.302 577.500 28.698 re W n
0.267 0.267 0.267 rg
BT 15.000 718.042 Td /F2 21.0 Tf  [(Human satellite cells: identification on human muscle fibres)] TJ ET
Q
0.271 0.267 0.267 rg
BT 15.000 700.036 Td /F3 9.8 Tf  [(January 19, 2012)] TJ ET
BT 26.250 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
BT 31.671 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 37.092 688.195 Td /F1 9.8 Tf  [(Luisa Boldrin)] TJ ET
0.271 0.267 0.267 rg
BT 93.447 688.195 Td /F1 9.8 Tf  [(, )] TJ ET
0.267 0.267 0.267 rg
BT 98.868 688.195 Td /F1 9.8 Tf  [(Jennifer E Morgan)] TJ ET
0.271 0.267 0.267 rg
BT 26.250 676.290 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE. Human satellite cells: identification on human muscle fibres. PLOS Currents Muscular Dystrophy. 2012 )] TJ ET
BT 26.250 664.386 Td /F1 9.8 Tf  [(Jan 19 . Edition 1. doi: 10.1371/currents.RRN1294.)] TJ ET
q
15.000 20.421 577.500 641.584 re W n
0.271 0.267 0.267 rg
BT 26.250 635.283 Td /F4 12.0 Tf  [(Abstract)] TJ ET
BT 26.250 615.329 Td /F1 9.8 Tf  [(Satellite cells, normally quiescent underneath the myofibre basal lamina, are skeletal muscle stem cells responsible for )] TJ ET
BT 26.250 603.424 Td /F1 9.8 Tf  [(postnatal muscle growth, repair and regeneration. Since their scarcity and small size have limited study on transverse muscle )] TJ ET
BT 26.250 591.519 Td /F1 9.8 Tf  [(sections, techniques to isolate individual myofibres, bearing their attendant satellite cells, were developed. Studies on mouse )] TJ ET
BT 26.250 579.615 Td /F1 9.8 Tf  [(myofibres have generated much information on satellite cells, but the limited availability and small size of human muscle )] TJ ET
BT 26.250 567.710 Td /F1 9.8 Tf  [(biopsies have hampered equivalent studies of satellite cells on human myofibres. Here, we identified satellite cells on fragments )] TJ ET
BT 26.250 555.805 Td /F1 9.8 Tf  [(of human and mouse myofibres, using a method applicable to small muscle biopsies.)] TJ ET
BT 26.250 519.203 Td /F4 12.0 Tf  [(Funding Statement)] TJ ET
BT 26.250 499.248 Td /F1 9.8 Tf  [(LB is founded by the Muscular Dystrophy Campaign \(MDC\) and JM holds a Wellcome Trust University award.)] TJ ET
BT 26.250 470.146 Td /F4 12.0 Tf  [(Introduction)] TJ ET
BT 26.250 450.192 Td /F1 9.8 Tf  [(Satellite cells are the principal stem cells of skeletal muscle, responsible for its homeostasis, maintenance, growth and )] TJ ET
BT 26.250 438.287 Td /F1 9.8 Tf  [(regeneration through adulthood)] TJ ET
0.267 0.267 0.267 rg
BT 162.292 438.287 Td /F1 9.8 Tf  [([1])] TJ ET
0.271 0.267 0.267 rg
BT 173.134 438.287 Td /F1 9.8 Tf  [( . Satellite cells are located in a niche between the basal lamina and sarcolemma of muscle )] TJ ET
BT 26.250 426.382 Td /F1 9.8 Tf  [(fibres )] TJ ET
0.267 0.267 0.267 rg
BT 52.799 426.382 Td /F1 9.8 Tf  [([2])] TJ ET
0.271 0.267 0.267 rg
BT 63.641 426.382 Td /F1 9.8 Tf  [( . Their rarity and small size are limitations for studying them on transverse muscle sections; therefore techniques were )] TJ ET
BT 26.250 414.477 Td /F1 9.8 Tf  [(developed to prepare isolated muscle fibres, bearing their attendant satellite cells, from rodent muscles)] TJ ET
0.267 0.267 0.267 rg
BT 469.524 414.477 Td /F1 9.8 Tf  [( [3] )] TJ ET
BT 485.787 414.477 Td /F1 9.8 Tf  [([4] )] TJ ET
0.271 0.267 0.267 rg
BT 499.339 414.477 Td /F1 9.8 Tf  [(. These isolated, )] TJ ET
BT 26.250 402.573 Td /F1 9.8 Tf  [(viable muscle fibres, particularly when prepared from genetically-modified mice in which either myonuclei, or satellite cells, )] TJ ET
BT 26.250 390.668 Td /F1 9.8 Tf  [(express a reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 390.668 Td /F1 9.8 Tf  [([5] [6] [7] ,)] TJ ET
0.271 0.267 0.267 rg
BT 175.834 390.668 Td /F1 9.8 Tf  [( have provided a model that has facilitated discovery of novel satellite cell markers, factors )] TJ ET
BT 26.250 378.763 Td /F1 9.8 Tf  [(controlling satellite cell quiescence, activation, proliferation, differentiation, self-renewal and determination of their in vivo )] TJ ET
BT 26.250 366.858 Td /F1 9.8 Tf  [(regenerative capacity )] TJ ET
0.267 0.267 0.267 rg
BT 121.624 366.858 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 132.466 366.858 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 347.454 Td /F1 9.8 Tf  [(Quiescent satellite cells on isolated mouse muscle fibres express Pax7 )] TJ ET
0.267 0.267 0.267 rg
BT 334.574 347.454 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 345.416 347.454 Td /F1 9.8 Tf  [( , M-cadherin )] TJ ET
0.267 0.267 0.267 rg
BT 405.018 347.454 Td /F1 9.8 Tf  [([10])] TJ ET
0.271 0.267 0.267 rg
BT 421.281 347.454 Td /F1 9.8 Tf  [( , CD34, Myf5 )] TJ ET
0.267 0.267 0.267 rg
BT 483.593 347.454 Td /F1 9.8 Tf  [([11] )] TJ ET
0.271 0.267 0.267 rg
BT 502.567 347.454 Td /F1 9.8 Tf  [(caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 549.708 347.454 Td /F1 9.8 Tf  [([12] )] TJ ET
BT 26.250 335.549 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 42.513 335.549 Td /F1 9.8 Tf  [( , calcitonin receptor )] TJ ET
0.267 0.267 0.267 rg
BT 132.466 335.549 Td /F1 9.8 Tf  [([13] [14] )] TJ ET
0.271 0.267 0.267 rg
BT 170.413 335.549 Td /F1 9.8 Tf  [(, nestin )] TJ ET
0.267 0.267 0.267 rg
BT 204.558 335.549 Td /F1 9.8 Tf  [([15] )] TJ ET
0.271 0.267 0.267 rg
BT 223.531 335.549 Td /F1 9.8 Tf  [(and ?7-integrin )] TJ ET
0.267 0.267 0.267 rg
BT 291.274 335.549 Td /F1 9.8 Tf  [([13] [16])] TJ ET
0.271 0.267 0.267 rg
BT 326.511 335.549 Td /F1 9.8 Tf  [( . During activation, myogenic regulatory factors, such as )] TJ ET
BT 26.250 323.644 Td /F1 9.8 Tf  [(MyoD and myogenin, are expressed, and satellite cells either self-renew or differentiate )] TJ ET
0.267 0.267 0.267 rg
BT 405.028 323.644 Td /F1 9.8 Tf  [([8] [17] [18] [19] [20])] TJ ET
0.271 0.267 0.267 rg
BT 491.764 323.644 Td /F1 9.8 Tf  [( . However, both in )] TJ ET
BT 26.250 311.739 Td /F1 9.8 Tf  [(vivo and in vitro studies have demonstrated that the population of mouse satellite cells is heterogeneous, some being more )] TJ ET
BT 26.250 299.835 Td /F1 9.8 Tf  [(�stem cell� like \(i.e. able to regenerate skeletal muscle and functionally reconstitute the satellite cell compartment\) than others )] TJ ET
0.267 0.267 0.267 rg
BT 26.250 287.930 Td /F1 9.8 Tf  [([21] [22] [23] [24])] TJ ET
0.271 0.267 0.267 rg
BT 99.433 287.930 Td /F1 9.8 Tf  [( \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 157.407 287.930 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 168.249 287.930 Td /F1 9.8 Tf  [(\).)] TJ ET
BT 26.250 268.525 Td /F1 9.8 Tf  [(Unfortunately, due to the difficulty in obtaining human muscle in a size and state adequate to allow the preparation of viable )] TJ ET
BT 26.250 256.620 Td /F1 9.8 Tf  [(fibres, there is a gap in our knowledge of human, compared to mouse, satellite cells \(reviewed in )] TJ ET
0.267 0.267 0.267 rg
BT 444.593 256.620 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 455.435 256.620 Td /F1 9.8 Tf  [( \). As a prelude to studies )] TJ ET
BT 26.250 244.716 Td /F1 9.8 Tf  [(on quiescent human satellite cells, we wished to determine antibodies that can be reliably used for their identification. To do )] TJ ET
BT 26.250 232.811 Td /F1 9.8 Tf  [(this, we exploited a technique to isolate fragments of human fibres from muscle biopsies of a normal individual and a Duchenne )] TJ ET
BT 26.250 220.906 Td /F1 9.8 Tf  [(muscular dystrophy \(DMD\) patient. As controls, myofibre fragments from mouse muscles were prepared by the same method. )] TJ ET
BT 26.250 209.001 Td /F1 9.8 Tf  [(We show that Pax7 and M-cadherin are reliable markers of not only mouse, but also human, satellite cells. In contrast, human )] TJ ET
BT 26.250 197.097 Td /F1 9.8 Tf  [(satellite cells are not stained by an antibody to CD34 that recognizes mouse satellite cells and only a subpopulation of human, )] TJ ET
BT 26.250 185.192 Td /F1 9.8 Tf  [(whereas all mouse, satellite cells express caveolin-1 )] TJ ET
0.267 0.267 0.267 rg
BT 254.907 185.192 Td /F1 9.8 Tf  [([13])] TJ ET
0.271 0.267 0.267 rg
BT 271.170 185.192 Td /F1 9.8 Tf  [(. Conversely, an antibody specific to human ?1 integrin \(CD29\) )] TJ ET
BT 26.250 173.287 Td /F1 9.8 Tf  [(identifies human, but not mouse, satellite cells. We therefore conclude that Pax7 marker defines the majority of not only mouse, )] TJ ET
BT 26.250 161.382 Td /F1 9.8 Tf  [(but also human, satellite cells.)] TJ ET
BT 26.250 124.780 Td /F4 12.0 Tf  [(Materials and Methods)] TJ ET
BT 26.250 104.826 Td /F4 9.8 Tf  [(Ethics)] TJ ET
BT 26.250 85.421 Td /F1 9.8 Tf  [(Tissue sampling was approved by the Institutional Ethics Committee in compliance with national guidelines regarding the use of )] TJ ET
BT 26.250 73.516 Td /F1 9.8 Tf  [(biopsy tissue for research. Patient tissues were collected after written informed consent. Mice were bred and experimental )] TJ ET
BT 26.250 61.611 Td /F1 9.8 Tf  [(procedures were carried out in the Biological Services Unit, University College London, in accordance with the Animals )] TJ ET
BT 26.250 49.707 Td /F1 9.8 Tf  [(\(Scientific Procedures\) Act 1986.)] TJ ET
Q
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(1)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
8 0 obj
<< /Type /Font
/Subtype /Type1
/Name /F1
/BaseFont /Helvetica
/Encoding /WinAnsiEncoding
>>
endobj
9 0 obj
<< /Type /Font
/Subtype /Type1
/Name /F2
/BaseFont /Times-Bold
/Encoding /WinAnsiEncoding
>>
endobj
10 0 obj
<< /Type /Font
/Subtype /Type1
/Name /F3
/BaseFont /Times-Italic
/Encoding /WinAnsiEncoding
>>
endobj
11 0 obj
<< /Type /Font
/Subtype /Type1
/Name /F4
/BaseFont /Helvetica-Bold
/Encoding /WinAnsiEncoding
>>
endobj
12 0 obj
<<
/Type /XObject
/Subtype /Image
/Width 500
/Height 52
/Filter /FlateDecode
/DecodeParms << /Predictor 15 /Colors 1 /Columns 500 /BitsPerComponent 8>>
/ColorSpace /DeviceGray
/BitsPerComponent 8
/Length 144>>
stream
x���1
0���'ݲ؎�"e����{��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��dz��A��tlM0\
endstream
endobj
13 0 obj
<<
/Type /XObject
/Subtype /Image
/Width 500
/Height 52
/SMask 12 0 R
/Filter /FlateDecode
/DecodeParms << /Predictor 15 /Colors 3 /Columns 500 /BitsPerComponent 8>>
/ColorSpace /DeviceRGB
/BitsPerComponent 8
/Length 4099>>
stream
x��ݙ��������,�`q�G�8��`G�
v�#�;3�
d�����U��g�gz���B 
��R�4�A0�0���}�
0�0��ŝa����0�	y���m۶�3d�an�a��ng��l6;��ʓa�����{����;��0���(Mӯ_�
!�<W~���EQ��"�0c��s��>I��d2�L��OFQ�u]׵"�m�N��$I�$�N�ܯ���d�����w�0̯�Y���=i�f��\.7_`�Z�Ŧ���rysV�0�'֞���=�t<���0�`���q��l�w�q�������K۶��aE���S���Y&C%�����p8(�
Eї/_�4�������'�����r9�ooor
�(
��X:Ɗ���D�������///�7â(���E��UUё������9�V����A����eY��ooo�E��jy*7F�h�v��TU%���rY.�A�um���C����	��c��`���,��<?���0t]WŨ�APE�ur>r����k�N'ϻ
ð�k������l��V4#��)M��Ȳ�,KwE�ah��т�N'㵨���(�B�1166Y��DQDy*�%��Q��A��
4^�r���(|*�a~[��.#O��a��y�4eY
!� ��k���PV���\EQ��ɘ?}�Ⱥc;�*q7j�� ʲ�e�ߣT�Έ� ���BY���Yzm��=�c<&j��y!
�EQ��
yR@
= �Ȋ��u��p0�2ʁR�e�4
F�w�a0.�UU�	��ծ���g�t�^��{}�Gܻ���H�40
ޝ�ȸB
��t]����aw�/�GIGH��K�=O�
�;����)%:��3��i��ÿ���,����W
��� x~~��-X.�H�#����g���|
�f��h��
���,��MA�f&<�y�I���4�B8N�'Aж��Sab��$�z��M�0���S�I�`�c#|��苑�{�޳�>��sB�a�8ݧ�~�����Շ8n� 5߻�|6�᚞;f�i�!
�p0$ [=\�]��DU�e��}y2��n�:ni�����u�Q�;��yt�(�	�:��
����deF�]��U��(�s��
	��?���Ɵ��|�J�׆τj�rã'P�U�'���k#�8�;��Go��g�?���a�X{�4_*O�u]�X�ӭ8*�/���F��>�l�e��M�m���ݡ��A�a���� Y��������.�1!9���|xosG�(c59
u���6w��q�7l+��|F�� ��a�
�h%P���a��;�N��(�H?�y�Tg��jr���L(�ή�lf4FOJ�Ye#�Ƚ`j�ʲLӔ�OD�Ɣ[Y�)O���*�*q�7�z��R�O��V-�wM�\�
+��a�XɘQ�uԷ��n˻�:�}���(Ċ
�]F�͟Rq���т�*)7~�*��'�@�ܯx� �Ý�G�Y&�cj?��u�s��J��9��zaoH�
b���T��S�ߘ�+
���eB
��eu6��o6���%�U4x��XX
0:}�3Z�a��eY��eQ���a����vS�]�s��f+~�
�"&��8�]/�Rj�ڑ+����a~�c[|̊"c8��� "r���ۡۨ����X�4׊�Q�G]�	c�?����,p�q���ʚ�1�UUs;�NM�(�G�#��Y���䯠�j��>��S�d��0\�|E�5/�f����:�B}�k�ݘ^�j`��K%gP��3�<:�� �06�؉�m��|޶-֧�vp���n� \?.�������3+ނ��hV��6I^��0̏�W�iE"��h�����ݸ�x<���;�Z��=9<t����gڶe}g���w=�:$�S=�
"����0�,C{��0��e�Xl6�{v툢ȧ�|�#�&�����;���h�v��^q��-�=!�?n��!��}>���*�s�"�!�4]����@4B�����R�)�i�ީ��R�s)���q=TǘEt9��v�k�l6�*!څ��~�Z�GڶU�`����˥�޶��l6C�$I�?>WI�ıg�l63�L���5.����'I�\.�S.�K�$ʅ�˥���}�X,&�ɟ��A�J$_�8<��~�ZM�S�l+e�X,��J�DI�T�j�R�
��q�X�Wa>3FK<�*�C�{򟣠A���Rv��P�ߴ���WH�NjO<�-h�ӆ�U�'rG��+iન8�E�Lkcl���"W)���^�b����*�B��3��E��4x�F��(=~P*$MS��P�r��9DQ$�G�6ų�(
�u���1*����{���i�4U��_�R�X4.{+��T3��^�k
>	��c�*�;>]9��&nP�;O���u/���k���C"���0>QTK�p�#�q�v]�hǺ���tRVH�#�2��@C�;����2r������\������|>�Nza��y�ˋE �zW/�����u�Rc��>C�t!�7�a��#~/�����tx���z�XL�SJûY�{ ��
��d6���J�	�0�%6}��d��� ���m��_��i���e��p�����ji�>
&
�%%o�(�������k2�=ġ�]��0|~~�5��)ò0��������-)DZM�b�!n�nY����r�l6�kw`>
�=T�y��"�����t���axͲL�u�	�!�]:�
Ak�O�X�V��.Mӧ�'!D��ooo�z��n�d�~}}��f���Y�A"��7���w����v��B�V˲4��ݔe�$�t:�MC��x<�q��y'����,���߾}C��m[wPO#i�*O<����\.�ǣ��WQ��r�D
��?�`b�r���{YU_^^l
|�e���r��5[�n��%GNO�߂o߾)l�ZA���3���0������[|��d���Wœ4�XW~�Y�
��6;�U�F[����
nC�p�����_� ����>�A�,W�R�X9l�c���e1g�������f�a�4���Qm�q�?)�Y���y`j�ݰ�l/BH�ri!D�����G���^0���uyQ>$���P7��Y�{Lg����;ж��'J�]���F�_������cY�8�v��@L9ގ���fq�����	Ӝ
j�����r��@p�巉;����?MS[�g<8�u2ު�'ۣ,��3�4�����E>��PU�z������"����b�sYi�����=���;�*�H�����p;˲��kF$����O8�0a����[Y�������<�=зQ��ￋ�X,���M=Bto'�6�� �f�Ql�OOO6��f�q���˗/ϓ�5�i
�٘�(��K|_�YU	
��x�Q=wp:���t4�c�w�؆O	g=�iZ���(����æ�5d_�v(���繾ob�4rA��9�b,��CV@=���6O6��;�a�+��徰l���Q¯TyO��[9h�7񠞻���|��x೤\�p�hq$��艊���������A�����>҇��,˪��A4N�Q�%�+W�0.˲�����|��*u]�WA��I�JP\�"��Spa�@�����b��ŝ6����~E���>�:�A�m�4eY�îg�����X��붳9�N�=E��<u�x��t�������w]G�������"w]��V��X�X�nqWUU)EK�T~"tcp��V�uy�#���Xw��
΍��<�tJ2��ʥ�[��jԔ��$�R����m��k+
Ɔ��k�]Bc���%2�U����1�O�0��s/0��e2Hqz�B6�*B��=l����q�\�Ɗ�և�3�L��5�0:׉{۶}ߏ��r��l̯w����;�0�qE<��r9�Ͱ���9@�r�?�a��W�۶%�4X]�TR������u�0��5��=���4Ma���3�0n����f3,���in�a���+���.�˯��0�p�+$�0��r���0����0�	aqg�����3�|BX��a>!,��0���	��
endstream
endobj
14 0 obj
<< /Type /Annot
/Subtype /Link
/A 15 0 R
/Border [0 0 0]
/H /I
/Rect [ 15.0000 716.1516 551.0880 736.9416 ]
>>
endobj
15 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/article/human-satellite-cells-identification-on-1q26mf55j8ydd-8/)
>>
endobj
16 0 obj
<< /Type /Annot
/Subtype /Link
/A 17 0 R
/Border [0 0 0]
/H /I
/Rect [ 37.0920 687.2933 93.4470 697.2140 ]
>>
endobj
17 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/luisaboldrin/)
>>
endobj
18 0 obj
<< /Type /Annot
/Subtype /Link
/A 19 0 R
/Border [0 0 0]
/H /I
/Rect [ 98.8680 687.2933 178.5255 697.2140 ]
>>
endobj
19 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/jenniferemorgan/)
>>
endobj
20 0 obj
<< /Type /Annot
/Subtype /Link
/A 21 0 R
/Border [0 0 0]
/H /I
/Rect [ 162.2918 437.3851 173.1338 447.3057 ]
>>
endobj
21 0 obj
<< /Type /Action
>>
endobj
22 0 obj
<< /Type /Annot
/Subtype /Link
/A 23 0 R
/Border [0 0 0]
/H /I
/Rect [ 52.7993 425.4803 63.6412 435.4010 ]
>>
endobj
23 0 obj
<< /Type /Action
>>
endobj
24 0 obj
<< /Type /Annot
/Subtype /Link
/A 25 0 R
/Border [0 0 0]
/H /I
/Rect [ 469.5240 413.5756 485.7870 423.4962 ]
>>
endobj
25 0 obj
<< /Type /Action
>>
endobj
26 0 obj
<< /Type /Annot
/Subtype /Link
/A 27 0 R
/Border [0 0 0]
/H /I
/Rect [ 485.7870 413.5756 499.3395 423.4962 ]
>>
endobj
27 0 obj
<< /Type /Action
>>
endobj
28 0 obj
<< /Type /Annot
/Subtype /Link
/A 29 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 389.7661 175.8345 399.6867 ]
>>
endobj
29 0 obj
<< /Type /Action
>>
endobj
30 0 obj
<< /Type /Annot
/Subtype /Link
/A 31 0 R
/Border [0 0 0]
/H /I
/Rect [ 121.6245 365.9566 132.4665 375.8772 ]
>>
endobj
31 0 obj
<< /Type /Action
>>
endobj
32 0 obj
<< /Type /Annot
/Subtype /Link
/A 33 0 R
/Border [0 0 0]
/H /I
/Rect [ 334.5743 346.5518 345.4162 356.4725 ]
>>
endobj
33 0 obj
<< /Type /Action
>>
endobj
34 0 obj
<< /Type /Annot
/Subtype /Link
/A 35 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0180 346.5518 421.2810 356.4725 ]
>>
endobj
35 0 obj
<< /Type /Action
>>
endobj
36 0 obj
<< /Type /Annot
/Subtype /Link
/A 37 0 R
/Border [0 0 0]
/H /I
/Rect [ 483.5932 346.5518 502.5667 356.4725 ]
>>
endobj
37 0 obj
<< /Type /Action
>>
endobj
38 0 obj
<< /Type /Annot
/Subtype /Link
/A 39 0 R
/Border [0 0 0]
/H /I
/Rect [ 549.7080 346.5518 568.6815 356.4725 ]
>>
endobj
39 0 obj
<< /Type /Action
>>
endobj
40 0 obj
<< /Type /Annot
/Subtype /Link
/A 41 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 334.6471 42.5130 344.5677 ]
>>
endobj
41 0 obj
<< /Type /Action
>>
endobj
42 0 obj
<< /Type /Annot
/Subtype /Link
/A 43 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 334.6471 170.4135 344.5677 ]
>>
endobj
43 0 obj
<< /Type /Action
>>
endobj
44 0 obj
<< /Type /Annot
/Subtype /Link
/A 45 0 R
/Border [0 0 0]
/H /I
/Rect [ 204.5580 334.6471 223.5315 344.5677 ]
>>
endobj
45 0 obj
<< /Type /Action
>>
endobj
46 0 obj
<< /Type /Annot
/Subtype /Link
/A 47 0 R
/Border [0 0 0]
/H /I
/Rect [ 291.2745 334.6471 326.5110 344.5677 ]
>>
endobj
47 0 obj
<< /Type /Action
>>
endobj
48 0 obj
<< /Type /Annot
/Subtype /Link
/A 49 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0278 322.7423 491.7638 332.6630 ]
>>
endobj
49 0 obj
<< /Type /Action
>>
endobj
50 0 obj
<< /Type /Annot
/Subtype /Link
/A 51 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 287.0281 99.4335 296.9487 ]
>>
endobj
51 0 obj
<< /Type /Action
>>
endobj
52 0 obj
<< /Type /Annot
/Subtype /Link
/A 53 0 R
/Border [0 0 0]
/H /I
/Rect [ 157.4070 287.0281 168.2490 296.9487 ]
>>
endobj
53 0 obj
<< /Type /Action
>>
endobj
54 0 obj
<< /Type /Annot
/Subtype /Link
/A 55 0 R
/Border [0 0 0]
/H /I
/Rect [ 444.5933 255.7186 455.4352 265.6392 ]
>>
endobj
55 0 obj
<< /Type /Action
>>
endobj
56 0 obj
<< /Type /Annot
/Subtype /Link
/A 57 0 R
/Border [0 0 0]
/H /I
/Rect [ 254.9070 184.2901 271.1700 194.2107 ]
>>
endobj
57 0 obj
<< /Type /Action
>>
endobj
58 0 obj
<< /Type /Annot
/Subtype /Link
/A 59 0 R
/Border [0 0 0]
/H /I
/Rect [ 15.0000 716.1516 551.0880 736.9416 ]
>>
endobj
59 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/article/human-satellite-cells-identification-on-1q26mf55j8ydd-8/)
>>
endobj
60 0 obj
<< /Type /Annot
/Subtype /Link
/A 61 0 R
/Border [0 0 0]
/H /I
/Rect [ 37.0920 687.2933 93.4470 697.2140 ]
>>
endobj
61 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/luisaboldrin/)
>>
endobj
62 0 obj
<< /Type /Annot
/Subtype /Link
/A 63 0 R
/Border [0 0 0]
/H /I
/Rect [ 98.8680 687.2933 178.5255 697.2140 ]
>>
endobj
63 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/jenniferemorgan/)
>>
endobj
64 0 obj
<< /Type /Annot
/Subtype /Link
/A 65 0 R
/Border [0 0 0]
/H /I
/Rect [ 162.2918 437.3851 173.1338 447.3057 ]
>>
endobj
65 0 obj
<< /Type /Action
>>
endobj
66 0 obj
<< /Type /Annot
/Subtype /Link
/A 67 0 R
/Border [0 0 0]
/H /I
/Rect [ 52.7993 425.4803 63.6412 435.4010 ]
>>
endobj
67 0 obj
<< /Type /Action
>>
endobj
68 0 obj
<< /Type /Annot
/Subtype /Link
/A 69 0 R
/Border [0 0 0]
/H /I
/Rect [ 469.5240 413.5756 485.7870 423.4962 ]
>>
endobj
69 0 obj
<< /Type /Action
>>
endobj
70 0 obj
<< /Type /Annot
/Subtype /Link
/A 71 0 R
/Border [0 0 0]
/H /I
/Rect [ 485.7870 413.5756 499.3395 423.4962 ]
>>
endobj
71 0 obj
<< /Type /Action
>>
endobj
72 0 obj
<< /Type /Annot
/Subtype /Link
/A 73 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 389.7661 175.8345 399.6867 ]
>>
endobj
73 0 obj
<< /Type /Action
>>
endobj
74 0 obj
<< /Type /Annot
/Subtype /Link
/A 75 0 R
/Border [0 0 0]
/H /I
/Rect [ 121.6245 365.9566 132.4665 375.8772 ]
>>
endobj
75 0 obj
<< /Type /Action
>>
endobj
76 0 obj
<< /Type /Annot
/Subtype /Link
/A 77 0 R
/Border [0 0 0]
/H /I
/Rect [ 334.5743 346.5518 345.4162 356.4725 ]
>>
endobj
77 0 obj
<< /Type /Action
>>
endobj
78 0 obj
<< /Type /Annot
/Subtype /Link
/A 79 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0180 346.5518 421.2810 356.4725 ]
>>
endobj
79 0 obj
<< /Type /Action
>>
endobj
80 0 obj
<< /Type /Annot
/Subtype /Link
/A 81 0 R
/Border [0 0 0]
/H /I
/Rect [ 483.5932 346.5518 502.5667 356.4725 ]
>>
endobj
81 0 obj
<< /Type /Action
>>
endobj
82 0 obj
<< /Type /Annot
/Subtype /Link
/A 83 0 R
/Border [0 0 0]
/H /I
/Rect [ 549.7080 346.5518 568.6815 356.4725 ]
>>
endobj
83 0 obj
<< /Type /Action
>>
endobj
84 0 obj
<< /Type /Annot
/Subtype /Link
/A 85 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 334.6471 42.5130 344.5677 ]
>>
endobj
85 0 obj
<< /Type /Action
>>
endobj
86 0 obj
<< /Type /Annot
/Subtype /Link
/A 87 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 334.6471 170.4135 344.5677 ]
>>
endobj
87 0 obj
<< /Type /Action
>>
endobj
88 0 obj
<< /Type /Annot
/Subtype /Link
/A 89 0 R
/Border [0 0 0]
/H /I
/Rect [ 204.5580 334.6471 223.5315 344.5677 ]
>>
endobj
89 0 obj
<< /Type /Action
>>
endobj
90 0 obj
<< /Type /Annot
/Subtype /Link
/A 91 0 R
/Border [0 0 0]
/H /I
/Rect [ 291.2745 334.6471 326.5110 344.5677 ]
>>
endobj
91 0 obj
<< /Type /Action
>>
endobj
92 0 obj
<< /Type /Annot
/Subtype /Link
/A 93 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0278 322.7423 491.7638 332.6630 ]
>>
endobj
93 0 obj
<< /Type /Action
>>
endobj
94 0 obj
<< /Type /Annot
/Subtype /Link
/A 95 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 287.0281 99.4335 296.9487 ]
>>
endobj
95 0 obj
<< /Type /Action
>>
endobj
96 0 obj
<< /Type /Annot
/Subtype /Link
/A 97 0 R
/Border [0 0 0]
/H /I
/Rect [ 157.4070 287.0281 168.2490 296.9487 ]
>>
endobj
97 0 obj
<< /Type /Action
>>
endobj
98 0 obj
<< /Type /Annot
/Subtype /Link
/A 99 0 R
/Border [0 0 0]
/H /I
/Rect [ 444.5933 255.7186 455.4352 265.6392 ]
>>
endobj
99 0 obj
<< /Type /Action
>>
endobj
100 0 obj
<< /Type /Annot
/Subtype /Link
/A 101 0 R
/Border [0 0 0]
/H /I
/Rect [ 254.9070 184.2901 271.1700 194.2107 ]
>>
endobj
101 0 obj
<< /Type /Action
>>
endobj
102 0 obj
<< /Type /Annot
/Subtype /Link
/A 103 0 R
/Border [0 0 0]
/H /I
/Rect [ 15.0000 716.1516 551.0880 736.9416 ]
>>
endobj
103 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/article/human-satellite-cells-identification-on-1q26mf55j8ydd-8/)
>>
endobj
104 0 obj
<< /Type /Annot
/Subtype /Link
/A 105 0 R
/Border [0 0 0]
/H /I
/Rect [ 37.0920 687.2933 93.4470 697.2140 ]
>>
endobj
105 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/luisaboldrin/)
>>
endobj
106 0 obj
<< /Type /Annot
/Subtype /Link
/A 107 0 R
/Border [0 0 0]
/H /I
/Rect [ 98.8680 687.2933 178.5255 697.2140 ]
>>
endobj
107 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/author/jenniferemorgan/)
>>
endobj
108 0 obj
<< /Type /Annot
/Subtype /Link
/A 109 0 R
/Border [0 0 0]
/H /I
/Rect [ 162.2918 437.3851 173.1338 447.3057 ]
>>
endobj
109 0 obj
<< /Type /Action
>>
endobj
110 0 obj
<< /Type /Annot
/Subtype /Link
/A 111 0 R
/Border [0 0 0]
/H /I
/Rect [ 52.7993 425.4803 63.6412 435.4010 ]
>>
endobj
111 0 obj
<< /Type /Action
>>
endobj
112 0 obj
<< /Type /Annot
/Subtype /Link
/A 113 0 R
/Border [0 0 0]
/H /I
/Rect [ 469.5240 413.5756 485.7870 423.4962 ]
>>
endobj
113 0 obj
<< /Type /Action
>>
endobj
114 0 obj
<< /Type /Annot
/Subtype /Link
/A 115 0 R
/Border [0 0 0]
/H /I
/Rect [ 485.7870 413.5756 499.3395 423.4962 ]
>>
endobj
115 0 obj
<< /Type /Action
>>
endobj
116 0 obj
<< /Type /Annot
/Subtype /Link
/A 117 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 389.7661 175.8345 399.6867 ]
>>
endobj
117 0 obj
<< /Type /Action
>>
endobj
118 0 obj
<< /Type /Annot
/Subtype /Link
/A 119 0 R
/Border [0 0 0]
/H /I
/Rect [ 121.6245 365.9566 132.4665 375.8772 ]
>>
endobj
119 0 obj
<< /Type /Action
>>
endobj
120 0 obj
<< /Type /Annot
/Subtype /Link
/A 121 0 R
/Border [0 0 0]
/H /I
/Rect [ 334.5743 346.5518 345.4162 356.4725 ]
>>
endobj
121 0 obj
<< /Type /Action
>>
endobj
122 0 obj
<< /Type /Annot
/Subtype /Link
/A 123 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0180 346.5518 421.2810 356.4725 ]
>>
endobj
123 0 obj
<< /Type /Action
>>
endobj
124 0 obj
<< /Type /Annot
/Subtype /Link
/A 125 0 R
/Border [0 0 0]
/H /I
/Rect [ 483.5932 346.5518 502.5667 356.4725 ]
>>
endobj
125 0 obj
<< /Type /Action
>>
endobj
126 0 obj
<< /Type /Annot
/Subtype /Link
/A 127 0 R
/Border [0 0 0]
/H /I
/Rect [ 549.7080 346.5518 568.6815 356.4725 ]
>>
endobj
127 0 obj
<< /Type /Action
>>
endobj
128 0 obj
<< /Type /Annot
/Subtype /Link
/A 129 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 334.6471 42.5130 344.5677 ]
>>
endobj
129 0 obj
<< /Type /Action
>>
endobj
130 0 obj
<< /Type /Annot
/Subtype /Link
/A 131 0 R
/Border [0 0 0]
/H /I
/Rect [ 132.4665 334.6471 170.4135 344.5677 ]
>>
endobj
131 0 obj
<< /Type /Action
>>
endobj
132 0 obj
<< /Type /Annot
/Subtype /Link
/A 133 0 R
/Border [0 0 0]
/H /I
/Rect [ 204.5580 334.6471 223.5315 344.5677 ]
>>
endobj
133 0 obj
<< /Type /Action
>>
endobj
134 0 obj
<< /Type /Annot
/Subtype /Link
/A 135 0 R
/Border [0 0 0]
/H /I
/Rect [ 291.2745 334.6471 326.5110 344.5677 ]
>>
endobj
135 0 obj
<< /Type /Action
>>
endobj
136 0 obj
<< /Type /Annot
/Subtype /Link
/A 137 0 R
/Border [0 0 0]
/H /I
/Rect [ 405.0278 322.7423 491.7638 332.6630 ]
>>
endobj
137 0 obj
<< /Type /Action
>>
endobj
138 0 obj
<< /Type /Annot
/Subtype /Link
/A 139 0 R
/Border [0 0 0]
/H /I
/Rect [ 26.2500 287.0281 99.4335 296.9487 ]
>>
endobj
139 0 obj
<< /Type /Action
>>
endobj
140 0 obj
<< /Type /Annot
/Subtype /Link
/A 141 0 R
/Border [0 0 0]
/H /I
/Rect [ 157.4070 287.0281 168.2490 296.9487 ]
>>
endobj
141 0 obj
<< /Type /Action
>>
endobj
142 0 obj
<< /Type /Annot
/Subtype /Link
/A 143 0 R
/Border [0 0 0]
/H /I
/Rect [ 444.5933 255.7186 455.4352 265.6392 ]
>>
endobj
143 0 obj
<< /Type /Action
>>
endobj
144 0 obj
<< /Type /Annot
/Subtype /Link
/A 145 0 R
/Border [0 0 0]
/H /I
/Rect [ 254.9070 184.2901 271.1700 194.2107 ]
>>
endobj
145 0 obj
<< /Type /Action
>>
endobj
146 0 obj
<< /Type /Page
/Parent 3 0 R
/Annots [ 149 0 R 151 0 R 153 0 R 155 0 R 157 0 R 159 0 R 161 0 R 163 0 R 166 0 R 168 0 R 170 0 R 172 0 R 174 0 R 176 0 R 178 0 R 180 0 R 182 0 R 184 0 R 186 0 R 188 0 R 190 0 R 192 0 R 194 0 R 196 0 R ]
/Contents 147 0 R
>>
endobj
147 0 obj
<<
/Length 21926 >>
stream

0.271 0.267 0.267 rg
q
15.000 55.717 577.500 721.283 re W n
0.271 0.267 0.267 rg
BT 26.250 767.476 Td /F4 9.8 Tf  [(Isolation of human and mouse single fibre fragments)] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Biopsies of paraspinal muscle of a healthy adolescent patient \(female, 14 year old\) and of )] TJ ET
BT 414.817 748.071 Td /F5 9.8 Tf  [(extensor digitorum brevis)] TJ ET
BT 523.724 748.071 Td /F1 9.8 Tf  [( \(EDB\) )] TJ ET
BT 26.250 736.167 Td /F1 9.8 Tf  [(muscle of a DMD adolescent patient \(male, 11 year old\) were collected and fixed in 4% paraformaldehyde \(PFA, Sigma\) at )] TJ ET
BT 26.250 724.262 Td /F1 9.8 Tf  [(room temperature for 15 minutes. After sequential washes with phosphate buffer saline \(PBS, Sigma\), the sample was )] TJ ET
BT 26.250 712.357 Td /F1 9.8 Tf  [(embedded at 4 )] TJ ET
BT 94.003 716.245 Td /F1 8.7 Tf  [(?)] TJ ET
BT 98.821 712.357 Td /F1 9.8 Tf  [( C, first in 30% glycerol/PBS \(Sigma\) overnight, and then sequentially in 50%, 80% glycerol/PBS and finally )] TJ ET
BT 26.250 700.452 Td /F1 9.8 Tf  [(absolute glycerol until the sample sank to the bottom of the vial. The sample was then kept in glycerol at 4 )] TJ ET
BT 485.260 704.341 Td /F1 8.7 Tf  [(?)] TJ ET
BT 490.079 700.452 Td /F1 9.8 Tf  [( C until dissection of )] TJ ET
BT 26.250 688.548 Td /F1 9.8 Tf  [(fibre fragments.)] TJ ET
BT 26.250 669.143 Td /F1 9.8 Tf  [(Under a stereomicroscope \(Leica\), fine forceps were used to dissect out fragments of fibres and exclude any other tissues, e.g. )] TJ ET
BT 26.250 657.238 Td /F1 9.8 Tf  [(connective tissue.)] TJ ET
BT 26.250 637.833 Td /F1 9.8 Tf  [(As a control, )] TJ ET
BT 83.151 637.833 Td /F5 9.8 Tf  [(tibialis anterior)] TJ ET
BT 146.000 637.833 Td /F1 9.8 Tf  [( \(TA\) muscles from 2 transgenic 3F- )] TJ ET
BT 304.203 637.833 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 331.298 637.833 Td /F1 9.8 Tf  [( -2E mice \(female, 2 month old\), in which myonuclei, but )] TJ ET
BT 26.250 625.929 Td /F1 9.8 Tf  [(not satellite cells express the )] TJ ET
BT 153.595 625.929 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 180.690 625.929 Td /F1 9.8 Tf  [( reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 244.640 625.929 Td /F1 9.8 Tf  [([25])] TJ ET
0.271 0.267 0.267 rg
BT 260.903 625.929 Td /F1 9.8 Tf  [(, were harvested, fixed and dissected in the same way as the human )] TJ ET
BT 26.250 614.024 Td /F1 9.8 Tf  [(muscle biopsies.)] TJ ET
BT 26.250 594.619 Td /F4 9.8 Tf  [(Immunohistochemistry of single fibre fragments)] TJ ET
BT 26.250 575.214 Td /F1 9.8 Tf  [(Dissected fibre fragments were washed in PBS at room temperature, permeabilized with 0.5% Triton X-100 \(Sigma\) for 6 )] TJ ET
BT 26.250 563.310 Td /F1 9.8 Tf  [(minutes, blocked for 30 minutes with 10% goat serum, and incubated overnight at 4 )] TJ ET
BT 388.804 567.198 Td /F1 8.7 Tf  [(?)] TJ ET
BT 393.622 563.310 Td /F1 9.8 Tf  [( C with the following primary antibodies: )] TJ ET
BT 26.250 551.405 Td /F1 9.8 Tf  [(mouse monoclonal anti-Pax7 \(Developmental Studies Hybridoma Bank, Iowa City, IA\), rabbit polyclonal anti-laminin \(Sigma\), )] TJ ET
BT 26.250 539.500 Td /F1 9.8 Tf  [(mouse monoclonal lamin A/C \(Vector Laboratories\), rabbit polyclonal caveolin-1 \(Santa Cruz Biotechnology, Inc.\), mouse )] TJ ET
BT 26.250 527.595 Td /F1 9.8 Tf  [(monoclonal M-cadherin)] TJ ET
0.267 0.267 0.267 rg
BT 127.572 527.595 Td /F1 9.8 Tf  [( [10])] TJ ET
0.271 0.267 0.267 rg
BT 146.546 527.595 Td /F1 9.8 Tf  [(, rat monoclonal anti mouse CD34 \(clone RAM34; PharMinogen\), mouse monoclonal anti human )] TJ ET
BT 26.250 515.691 Td /F1 9.8 Tf  [(CD29 \(AbD Serotec\). Secondary antibodies used were: Alexa Fluor 488-conjugated goat anti-mouse Ig \(Molecular Probes\) and )] TJ ET
BT 26.250 503.786 Td /F1 9.8 Tf  [(Alexa Fluor 594-conjugated goat anti-rabbit Ig \(Molecular Probes\). Mouse fibre fragments were X-Gal stained as previously )] TJ ET
BT 26.250 491.881 Td /F1 9.8 Tf  [(described)] TJ ET
0.267 0.267 0.267 rg
BT 68.516 491.881 Td /F1 9.8 Tf  [( [11])] TJ ET
0.271 0.267 0.267 rg
BT 87.490 491.881 Td /F1 9.8 Tf  [(. Nuclei were counterstained with DAPI and mounted on slide in fluorescent mounting medium \(Dako\).)] TJ ET
BT 26.250 472.476 Td /F4 9.8 Tf  [(Microscopy)] TJ ET
BT 26.250 453.072 Td /F1 9.8 Tf  [(Fluorescence detection, bright-field microscopy and image capturing were done using a Zeiss Axiophot microscope \(Carl Zeiss\), )] TJ ET
BT 26.250 441.167 Td /F1 9.8 Tf  [(and Metamorph software \(Metamorph Productions\).)] TJ ET
BT 26.250 421.762 Td /F4 9.8 Tf  [(Data Analysis)] TJ ET
BT 26.250 402.357 Td /F1 9.8 Tf  [(Fibre length was calculated by using SigmaScan software in order to allow satellite cell quantification per 100�m fibre length.)] TJ ET
BT 26.250 365.755 Td /F4 12.0 Tf  [(Results)] TJ ET
BT 26.250 345.801 Td /F4 9.8 Tf  [(A high number of satellite cells is present in human fibres)] TJ ET
BT 26.250 326.396 Td /F1 9.8 Tf  [(All nuclei \(both myonuclei and satellite cell nuclei\) on isolated fibres harvested from normal human paraspinal muscle biopsies )] TJ ET
BT 26.250 314.491 Td /F1 9.8 Tf  [(expressed, as expected, human-specific lamin A/C in their nuclear membrane )] TJ ET
0.267 0.267 0.267 rg
BT 363.834 314.491 Td /F1 9.8 Tf  [([26] [27])] TJ ET
0.271 0.267 0.267 rg
BT 399.070 314.491 Td /F1 9.8 Tf  [(, confirming that the fixation and fibre )] TJ ET
BT 26.250 302.586 Td /F1 9.8 Tf  [(preparation did not interfere with immunostaining in human fibres \(Figure 1A-I\).)] TJ ET
BT 26.250 283.182 Td /F1 9.8 Tf  [(Pax7 antibody clearly marked both human \(Figure 1A II -III and Figure 2B\) and mouse \(Figure 2A\) satellite cells; co-staining )] TJ ET
BT 26.250 271.277 Td /F1 9.8 Tf  [(with laminin antibody confirmed that satellite cells were retained under the basal lamina of human fibres following the )] TJ ET
BT 26.250 259.372 Td /F1 9.8 Tf  [(preparation procedure \(Figure 1A- III\). The mean number of human Pax7+ve satellite cells per 100�m length of fibre was 0.9 )] TJ ET
BT 26.250 247.467 Td /F1 9.8 Tf  [(\(�0.05 SEM\) \(Figure 1B\). Considering that the paraspinal muscle is composed of fascicles of different lengths, )] TJ ET
0.267 0.267 0.267 rg
BT 503.386 247.467 Td /F1 9.8 Tf  [([28] )] TJ ET
0.271 0.267 0.267 rg
BT 522.359 247.467 Td /F1 9.8 Tf  [(it not )] TJ ET
BT 26.250 235.563 Td /F1 9.8 Tf  [(possible to determine the number of satellite cells per fibre. However, knowing that the mean number of satellite cells per TA )] TJ ET
BT 26.250 223.658 Td /F1 9.8 Tf  [(myofibre is 10)] TJ ET
0.267 0.267 0.267 rg
BT 86.934 223.658 Td /F1 9.8 Tf  [( [21])] TJ ET
0.271 0.267 0.267 rg
BT 105.907 223.658 Td /F1 9.8 Tf  [( and that the mean length of a TA myofibre is 15mm)] TJ ET
0.267 0.267 0.267 rg
BT 330.265 223.658 Td /F1 9.8 Tf  [( [29])] TJ ET
0.271 0.267 0.267 rg
BT 349.238 223.658 Td /F1 9.8 Tf  [(, we conclude that the density of satellite cells per )] TJ ET
BT 26.250 211.753 Td /F1 9.8 Tf  [(fibre length is higher in human paraspinal muscles than in mouse TA muscles.)] TJ ET
0.965 0.965 0.965 rg
26.250 55.717 555.000 146.155 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 201.872 m 581.250 201.872 l 581.250 201.122 l 26.250 201.122 l  f
q
112.500 0 0 112.500 35.250 79.622 cm /I3 Do
Q 
q
35.250 55.717 537.000 17.905 re W n
Q
Q
q
15.000 55.717 577.500 721.283 re W n
0.271 0.267 0.267 rg
BT 26.250 767.476 Td /F4 9.8 Tf  [(Isolation of human and mouse single fibre fragments)] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Biopsies of paraspinal muscle of a healthy adolescent patient \(female, 14 year old\) and of )] TJ ET
BT 414.817 748.071 Td /F5 9.8 Tf  [(extensor digitorum brevis)] TJ ET
BT 523.724 748.071 Td /F1 9.8 Tf  [( \(EDB\) )] TJ ET
BT 26.250 736.167 Td /F1 9.8 Tf  [(muscle of a DMD adolescent patient \(male, 11 year old\) were collected and fixed in 4% paraformaldehyde \(PFA, Sigma\) at )] TJ ET
BT 26.250 724.262 Td /F1 9.8 Tf  [(room temperature for 15 minutes. After sequential washes with phosphate buffer saline \(PBS, Sigma\), the sample was )] TJ ET
BT 26.250 712.357 Td /F1 9.8 Tf  [(embedded at 4 )] TJ ET
BT 94.003 716.245 Td /F1 8.7 Tf  [(?)] TJ ET
BT 98.821 712.357 Td /F1 9.8 Tf  [( C, first in 30% glycerol/PBS \(Sigma\) overnight, and then sequentially in 50%, 80% glycerol/PBS and finally )] TJ ET
BT 26.250 700.452 Td /F1 9.8 Tf  [(absolute glycerol until the sample sank to the bottom of the vial. The sample was then kept in glycerol at 4 )] TJ ET
BT 485.260 704.341 Td /F1 8.7 Tf  [(?)] TJ ET
BT 490.079 700.452 Td /F1 9.8 Tf  [( C until dissection of )] TJ ET
BT 26.250 688.548 Td /F1 9.8 Tf  [(fibre fragments.)] TJ ET
BT 26.250 669.143 Td /F1 9.8 Tf  [(Under a stereomicroscope \(Leica\), fine forceps were used to dissect out fragments of fibres and exclude any other tissues, e.g. )] TJ ET
BT 26.250 657.238 Td /F1 9.8 Tf  [(connective tissue.)] TJ ET
BT 26.250 637.833 Td /F1 9.8 Tf  [(As a control, )] TJ ET
BT 83.151 637.833 Td /F5 9.8 Tf  [(tibialis anterior)] TJ ET
BT 146.000 637.833 Td /F1 9.8 Tf  [( \(TA\) muscles from 2 transgenic 3F- )] TJ ET
BT 304.203 637.833 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 331.298 637.833 Td /F1 9.8 Tf  [( -2E mice \(female, 2 month old\), in which myonuclei, but )] TJ ET
BT 26.250 625.929 Td /F1 9.8 Tf  [(not satellite cells express the )] TJ ET
BT 153.595 625.929 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 180.690 625.929 Td /F1 9.8 Tf  [( reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 244.640 625.929 Td /F1 9.8 Tf  [([25])] TJ ET
0.271 0.267 0.267 rg
BT 260.903 625.929 Td /F1 9.8 Tf  [(, were harvested, fixed and dissected in the same way as the human )] TJ ET
BT 26.250 614.024 Td /F1 9.8 Tf  [(muscle biopsies.)] TJ ET
BT 26.250 594.619 Td /F4 9.8 Tf  [(Immunohistochemistry of single fibre fragments)] TJ ET
BT 26.250 575.214 Td /F1 9.8 Tf  [(Dissected fibre fragments were washed in PBS at room temperature, permeabilized with 0.5% Triton X-100 \(Sigma\) for 6 )] TJ ET
BT 26.250 563.310 Td /F1 9.8 Tf  [(minutes, blocked for 30 minutes with 10% goat serum, and incubated overnight at 4 )] TJ ET
BT 388.804 567.198 Td /F1 8.7 Tf  [(?)] TJ ET
BT 393.622 563.310 Td /F1 9.8 Tf  [( C with the following primary antibodies: )] TJ ET
BT 26.250 551.405 Td /F1 9.8 Tf  [(mouse monoclonal anti-Pax7 \(Developmental Studies Hybridoma Bank, Iowa City, IA\), rabbit polyclonal anti-laminin \(Sigma\), )] TJ ET
BT 26.250 539.500 Td /F1 9.8 Tf  [(mouse monoclonal lamin A/C \(Vector Laboratories\), rabbit polyclonal caveolin-1 \(Santa Cruz Biotechnology, Inc.\), mouse )] TJ ET
BT 26.250 527.595 Td /F1 9.8 Tf  [(monoclonal M-cadherin)] TJ ET
0.267 0.267 0.267 rg
BT 127.572 527.595 Td /F1 9.8 Tf  [( [10])] TJ ET
0.271 0.267 0.267 rg
BT 146.546 527.595 Td /F1 9.8 Tf  [(, rat monoclonal anti mouse CD34 \(clone RAM34; PharMinogen\), mouse monoclonal anti human )] TJ ET
BT 26.250 515.691 Td /F1 9.8 Tf  [(CD29 \(AbD Serotec\). Secondary antibodies used were: Alexa Fluor 488-conjugated goat anti-mouse Ig \(Molecular Probes\) and )] TJ ET
BT 26.250 503.786 Td /F1 9.8 Tf  [(Alexa Fluor 594-conjugated goat anti-rabbit Ig \(Molecular Probes\). Mouse fibre fragments were X-Gal stained as previously )] TJ ET
BT 26.250 491.881 Td /F1 9.8 Tf  [(described)] TJ ET
0.267 0.267 0.267 rg
BT 68.516 491.881 Td /F1 9.8 Tf  [( [11])] TJ ET
0.271 0.267 0.267 rg
BT 87.490 491.881 Td /F1 9.8 Tf  [(. Nuclei were counterstained with DAPI and mounted on slide in fluorescent mounting medium \(Dako\).)] TJ ET
BT 26.250 472.476 Td /F4 9.8 Tf  [(Microscopy)] TJ ET
BT 26.250 453.072 Td /F1 9.8 Tf  [(Fluorescence detection, bright-field microscopy and image capturing were done using a Zeiss Axiophot microscope \(Carl Zeiss\), )] TJ ET
BT 26.250 441.167 Td /F1 9.8 Tf  [(and Metamorph software \(Metamorph Productions\).)] TJ ET
BT 26.250 421.762 Td /F4 9.8 Tf  [(Data Analysis)] TJ ET
BT 26.250 402.357 Td /F1 9.8 Tf  [(Fibre length was calculated by using SigmaScan software in order to allow satellite cell quantification per 100�m fibre length.)] TJ ET
BT 26.250 365.755 Td /F4 12.0 Tf  [(Results)] TJ ET
BT 26.250 345.801 Td /F4 9.8 Tf  [(A high number of satellite cells is present in human fibres)] TJ ET
BT 26.250 326.396 Td /F1 9.8 Tf  [(All nuclei \(both myonuclei and satellite cell nuclei\) on isolated fibres harvested from normal human paraspinal muscle biopsies )] TJ ET
BT 26.250 314.491 Td /F1 9.8 Tf  [(expressed, as expected, human-specific lamin A/C in their nuclear membrane )] TJ ET
0.267 0.267 0.267 rg
BT 363.834 314.491 Td /F1 9.8 Tf  [([26] [27])] TJ ET
0.271 0.267 0.267 rg
BT 399.070 314.491 Td /F1 9.8 Tf  [(, confirming that the fixation and fibre )] TJ ET
BT 26.250 302.586 Td /F1 9.8 Tf  [(preparation did not interfere with immunostaining in human fibres \(Figure 1A-I\).)] TJ ET
BT 26.250 283.182 Td /F1 9.8 Tf  [(Pax7 antibody clearly marked both human \(Figure 1A II -III and Figure 2B\) and mouse \(Figure 2A\) satellite cells; co-staining )] TJ ET
BT 26.250 271.277 Td /F1 9.8 Tf  [(with laminin antibody confirmed that satellite cells were retained under the basal lamina of human fibres following the )] TJ ET
BT 26.250 259.372 Td /F1 9.8 Tf  [(preparation procedure \(Figure 1A- III\). The mean number of human Pax7+ve satellite cells per 100�m length of fibre was 0.9 )] TJ ET
BT 26.250 247.467 Td /F1 9.8 Tf  [(\(�0.05 SEM\) \(Figure 1B\). Considering that the paraspinal muscle is composed of fascicles of different lengths, )] TJ ET
0.267 0.267 0.267 rg
BT 503.386 247.467 Td /F1 9.8 Tf  [([28] )] TJ ET
0.271 0.267 0.267 rg
BT 522.359 247.467 Td /F1 9.8 Tf  [(it not )] TJ ET
BT 26.250 235.563 Td /F1 9.8 Tf  [(possible to determine the number of satellite cells per fibre. However, knowing that the mean number of satellite cells per TA )] TJ ET
BT 26.250 223.658 Td /F1 9.8 Tf  [(myofibre is 10)] TJ ET
0.267 0.267 0.267 rg
BT 86.934 223.658 Td /F1 9.8 Tf  [( [21])] TJ ET
0.271 0.267 0.267 rg
BT 105.907 223.658 Td /F1 9.8 Tf  [( and that the mean length of a TA myofibre is 15mm)] TJ ET
0.267 0.267 0.267 rg
BT 330.265 223.658 Td /F1 9.8 Tf  [( [29])] TJ ET
0.271 0.267 0.267 rg
BT 349.238 223.658 Td /F1 9.8 Tf  [(, we conclude that the density of satellite cells per )] TJ ET
BT 26.250 211.753 Td /F1 9.8 Tf  [(fibre length is higher in human paraspinal muscles than in mouse TA muscles.)] TJ ET
0.965 0.965 0.965 rg
26.250 55.717 555.000 146.155 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 201.872 m 581.250 201.872 l 581.250 201.122 l 26.250 201.122 l  f
q
112.500 0 0 112.500 35.250 79.622 cm /I3 Do
Q 
q
35.250 55.717 537.000 17.905 re W n
Q
Q
q
15.000 55.717 577.500 721.283 re W n
0.271 0.267 0.267 rg
BT 26.250 767.476 Td /F4 9.8 Tf  [(Isolation of human and mouse single fibre fragments)] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Biopsies of paraspinal muscle of a healthy adolescent patient \(female, 14 year old\) and of )] TJ ET
BT 414.817 748.071 Td /F5 9.8 Tf  [(extensor digitorum brevis)] TJ ET
BT 523.724 748.071 Td /F1 9.8 Tf  [( \(EDB\) )] TJ ET
BT 26.250 736.167 Td /F1 9.8 Tf  [(muscle of a DMD adolescent patient \(male, 11 year old\) were collected and fixed in 4% paraformaldehyde \(PFA, Sigma\) at )] TJ ET
BT 26.250 724.262 Td /F1 9.8 Tf  [(room temperature for 15 minutes. After sequential washes with phosphate buffer saline \(PBS, Sigma\), the sample was )] TJ ET
BT 26.250 712.357 Td /F1 9.8 Tf  [(embedded at 4 )] TJ ET
BT 94.003 716.245 Td /F1 8.7 Tf  [(?)] TJ ET
BT 98.821 712.357 Td /F1 9.8 Tf  [( C, first in 30% glycerol/PBS \(Sigma\) overnight, and then sequentially in 50%, 80% glycerol/PBS and finally )] TJ ET
BT 26.250 700.452 Td /F1 9.8 Tf  [(absolute glycerol until the sample sank to the bottom of the vial. The sample was then kept in glycerol at 4 )] TJ ET
BT 485.260 704.341 Td /F1 8.7 Tf  [(?)] TJ ET
BT 490.079 700.452 Td /F1 9.8 Tf  [( C until dissection of )] TJ ET
BT 26.250 688.548 Td /F1 9.8 Tf  [(fibre fragments.)] TJ ET
BT 26.250 669.143 Td /F1 9.8 Tf  [(Under a stereomicroscope \(Leica\), fine forceps were used to dissect out fragments of fibres and exclude any other tissues, e.g. )] TJ ET
BT 26.250 657.238 Td /F1 9.8 Tf  [(connective tissue.)] TJ ET
BT 26.250 637.833 Td /F1 9.8 Tf  [(As a control, )] TJ ET
BT 83.151 637.833 Td /F5 9.8 Tf  [(tibialis anterior)] TJ ET
BT 146.000 637.833 Td /F1 9.8 Tf  [( \(TA\) muscles from 2 transgenic 3F- )] TJ ET
BT 304.203 637.833 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 331.298 637.833 Td /F1 9.8 Tf  [( -2E mice \(female, 2 month old\), in which myonuclei, but )] TJ ET
BT 26.250 625.929 Td /F1 9.8 Tf  [(not satellite cells express the )] TJ ET
BT 153.595 625.929 Td /F5 9.8 Tf  [(nLacZ)] TJ ET
BT 180.690 625.929 Td /F1 9.8 Tf  [( reporter gene )] TJ ET
0.267 0.267 0.267 rg
BT 244.640 625.929 Td /F1 9.8 Tf  [([25])] TJ ET
0.271 0.267 0.267 rg
BT 260.903 625.929 Td /F1 9.8 Tf  [(, were harvested, fixed and dissected in the same way as the human )] TJ ET
BT 26.250 614.024 Td /F1 9.8 Tf  [(muscle biopsies.)] TJ ET
BT 26.250 594.619 Td /F4 9.8 Tf  [(Immunohistochemistry of single fibre fragments)] TJ ET
BT 26.250 575.214 Td /F1 9.8 Tf  [(Dissected fibre fragments were washed in PBS at room temperature, permeabilized with 0.5% Triton X-100 \(Sigma\) for 6 )] TJ ET
BT 26.250 563.310 Td /F1 9.8 Tf  [(minutes, blocked for 30 minutes with 10% goat serum, and incubated overnight at 4 )] TJ ET
BT 388.804 567.198 Td /F1 8.7 Tf  [(?)] TJ ET
BT 393.622 563.310 Td /F1 9.8 Tf  [( C with the following primary antibodies: )] TJ ET
BT 26.250 551.405 Td /F1 9.8 Tf  [(mouse monoclonal anti-Pax7 \(Developmental Studies Hybridoma Bank, Iowa City, IA\), rabbit polyclonal anti-laminin \(Sigma\), )] TJ ET
BT 26.250 539.500 Td /F1 9.8 Tf  [(mouse monoclonal lamin A/C \(Vector Laboratories\), rabbit polyclonal caveolin-1 \(Santa Cruz Biotechnology, Inc.\), mouse )] TJ ET
BT 26.250 527.595 Td /F1 9.8 Tf  [(monoclonal M-cadherin)] TJ ET
0.267 0.267 0.267 rg
BT 127.572 527.595 Td /F1 9.8 Tf  [( [10])] TJ ET
0.271 0.267 0.267 rg
BT 146.546 527.595 Td /F1 9.8 Tf  [(, rat monoclonal anti mouse CD34 \(clone RAM34; PharMinogen\), mouse monoclonal anti human )] TJ ET
BT 26.250 515.691 Td /F1 9.8 Tf  [(CD29 \(AbD Serotec\). Secondary antibodies used were: Alexa Fluor 488-conjugated goat anti-mouse Ig \(Molecular Probes\) and )] TJ ET
BT 26.250 503.786 Td /F1 9.8 Tf  [(Alexa Fluor 594-conjugated goat anti-rabbit Ig \(Molecular Probes\). Mouse fibre fragments were X-Gal stained as previously )] TJ ET
BT 26.250 491.881 Td /F1 9.8 Tf  [(described)] TJ ET
0.267 0.267 0.267 rg
BT 68.516 491.881 Td /F1 9.8 Tf  [( [11])] TJ ET
0.271 0.267 0.267 rg
BT 87.490 491.881 Td /F1 9.8 Tf  [(. Nuclei were counterstained with DAPI and mounted on slide in fluorescent mounting medium \(Dako\).)] TJ ET
BT 26.250 472.476 Td /F4 9.8 Tf  [(Microscopy)] TJ ET
BT 26.250 453.072 Td /F1 9.8 Tf  [(Fluorescence detection, bright-field microscopy and image capturing were done using a Zeiss Axiophot microscope \(Carl Zeiss\), )] TJ ET
BT 26.250 441.167 Td /F1 9.8 Tf  [(and Metamorph software \(Metamorph Productions\).)] TJ ET
BT 26.250 421.762 Td /F4 9.8 Tf  [(Data Analysis)] TJ ET
BT 26.250 402.357 Td /F1 9.8 Tf  [(Fibre length was calculated by using SigmaScan software in order to allow satellite cell quantification per 100�m fibre length.)] TJ ET
BT 26.250 365.755 Td /F4 12.0 Tf  [(Results)] TJ ET
BT 26.250 345.801 Td /F4 9.8 Tf  [(A high number of satellite cells is present in human fibres)] TJ ET
BT 26.250 326.396 Td /F1 9.8 Tf  [(All nuclei \(both myonuclei and satellite cell nuclei\) on isolated fibres harvested from normal human paraspinal muscle biopsies )] TJ ET
BT 26.250 314.491 Td /F1 9.8 Tf  [(expressed, as expected, human-specific lamin A/C in their nuclear membrane )] TJ ET
0.267 0.267 0.267 rg
BT 363.834 314.491 Td /F1 9.8 Tf  [([26] [27])] TJ ET
0.271 0.267 0.267 rg
BT 399.070 314.491 Td /F1 9.8 Tf  [(, confirming that the fixation and fibre )] TJ ET
BT 26.250 302.586 Td /F1 9.8 Tf  [(preparation did not interfere with immunostaining in human fibres \(Figure 1A-I\).)] TJ ET
BT 26.250 283.182 Td /F1 9.8 Tf  [(Pax7 antibody clearly marked both human \(Figure 1A II -III and Figure 2B\) and mouse \(Figure 2A\) satellite cells; co-staining )] TJ ET
BT 26.250 271.277 Td /F1 9.8 Tf  [(with laminin antibody confirmed that satellite cells were retained under the basal lamina of human fibres following the )] TJ ET
BT 26.250 259.372 Td /F1 9.8 Tf  [(preparation procedure \(Figure 1A- III\). The mean number of human Pax7+ve satellite cells per 100�m length of fibre was 0.9 )] TJ ET
BT 26.250 247.467 Td /F1 9.8 Tf  [(\(�0.05 SEM\) \(Figure 1B\). Considering that the paraspinal muscle is composed of fascicles of different lengths, )] TJ ET
0.267 0.267 0.267 rg
BT 503.386 247.467 Td /F1 9.8 Tf  [([28] )] TJ ET
0.271 0.267 0.267 rg
BT 522.359 247.467 Td /F1 9.8 Tf  [(it not )] TJ ET
BT 26.250 235.563 Td /F1 9.8 Tf  [(possible to determine the number of satellite cells per fibre. However, knowing that the mean number of satellite cells per TA )] TJ ET
BT 26.250 223.658 Td /F1 9.8 Tf  [(myofibre is 10)] TJ ET
0.267 0.267 0.267 rg
BT 86.934 223.658 Td /F1 9.8 Tf  [( [21])] TJ ET
0.271 0.267 0.267 rg
BT 105.907 223.658 Td /F1 9.8 Tf  [( and that the mean length of a TA myofibre is 15mm)] TJ ET
0.267 0.267 0.267 rg
BT 330.265 223.658 Td /F1 9.8 Tf  [( [29])] TJ ET
0.271 0.267 0.267 rg
BT 349.238 223.658 Td /F1 9.8 Tf  [(, we conclude that the density of satellite cells per )] TJ ET
BT 26.250 211.753 Td /F1 9.8 Tf  [(fibre length is higher in human paraspinal muscles than in mouse TA muscles.)] TJ ET
0.965 0.965 0.965 rg
26.250 55.717 555.000 146.155 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 201.872 m 581.250 201.872 l 581.250 201.122 l 26.250 201.122 l  f
q
112.500 0 0 112.500 35.250 79.622 cm /I3 Do
Q 
q
35.250 55.717 537.000 17.905 re W n
Q
Q
q
112.500 0 0 112.500 35.250 79.622 cm /I3 Do
Q 
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(2)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
148 0 obj
<< /Type /Font
/Subtype /Type1
/Name /F5
/BaseFont /Helvetica-Oblique
/Encoding /WinAnsiEncoding
>>
endobj
149 0 obj
<< /Type /Annot
/Subtype /Link
/A 150 0 R
/Border [0 0 0]
/H /I
/Rect [ 244.6402 625.0268 260.9032 634.9474 ]
>>
endobj
150 0 obj
<< /Type /Action
>>
endobj
151 0 obj
<< /Type /Annot
/Subtype /Link
/A 152 0 R
/Border [0 0 0]
/H /I
/Rect [ 127.5720 526.6936 146.5455 536.6142 ]
>>
endobj
152 0 obj
<< /Type /Action
>>
endobj
153 0 obj
<< /Type /Annot
/Subtype /Link
/A 154 0 R
/Border [0 0 0]
/H /I
/Rect [ 68.5162 490.9793 87.4898 500.8999 ]
>>
endobj
154 0 obj
<< /Type /Action
>>
endobj
155 0 obj
<< /Type /Annot
/Subtype /Link
/A 156 0 R
/Border [0 0 0]
/H /I
/Rect [ 363.8340 313.5893 399.0705 323.5100 ]
>>
endobj
156 0 obj
<< /Type /Action
>>
endobj
157 0 obj
<< /Type /Annot
/Subtype /Link
/A 158 0 R
/Border [0 0 0]
/H /I
/Rect [ 503.3857 246.5656 522.3592 256.4862 ]
>>
endobj
158 0 obj
<< /Type /Action
>>
endobj
159 0 obj
<< /Type /Annot
/Subtype /Link
/A 160 0 R
/Border [0 0 0]
/H /I
/Rect [ 86.9340 222.7561 105.9075 232.6767 ]
>>
endobj
160 0 obj
<< /Type /Action
>>
endobj
161 0 obj
<< /Type /Annot
/Subtype /Link
/A 162 0 R
/Border [0 0 0]
/H /I
/Rect [ 330.2647 222.7561 349.2382 232.6767 ]
>>
endobj
162 0 obj
<< /Type /Action
>>
endobj
163 0 obj
<< /Type /Annot
/Subtype /Link
/A 164 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 79.6223 147.7500 192.1223 ]
>>
endobj
164 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-1-new.jpg)
>>
endobj
165 0 obj
<<
/Type /XObject
/Subtype /Image
/Width 150
/Height 150
/ColorSpace /DeviceRGB
/Filter /DCTDecode
/BitsPerComponent 8
/Length 6678>>
stream
����JFIF



��;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90
��C




��C
		

����
"

��







	
���

}
!1AQa"q2���#B��R��$3br�	
%&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz���������������������������������������������������������������������������










	
���

w
!1AQaq"2�B����	#3R�br�
$4�%�&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz��������������������������������������������������������������������������
?���7�>o�˼�ԯ�U���j����Я�-�������t�����������+��9�9D�����Ҭ����xj�`v�R=*��(���|��ߎkb�Qk�i،��z�ov���$�W�Z�����n-)(�~��Ү��� wZ�A`��R	�����+>��P^�U2}@�WD1���<�O	O��;�3�>>m�����gw�<S��Ek�� H�Hu}+�_�F�����s���}����N�29# ��{���$��/n����_�?>�NT�J��M���M��6����!Q��[#�`՘�˕�4Q��ym����<�P�����br	���E�a�o��V�{oek^+W{-.�s_FAE=YZBd�����	�1���w����L��bk���r'��q�mǯ�+洷����9U�	��<arm�������z����2j���,��
�k����J�����on���>�.�a��T���k7�a�FAW@��‘���d���9��
l�:�j*9m�F<�m����W����o�I�2��LZ%Jt�j�o�}U�n�G�����s�!��q�O�|�>��n��2��U&R�
�C^��]���+�X��T��=��#�vz9�5�K}u����G	9E�;�3�g���&�����?�Ί���7��_���a����ם���W|d������RT�+5�������q��w�?��+�?���E�/�o��x��7�)�f��鴭*��&��
IL)�I$rg�n�G8�d_��xS�r�6?�2�?�����ӎ�-��-
!
�ڗk_����Ϟ���.Np=��Y��.m�h}ɜ���׾��5�O���I�a�\��/X�MP��c�3��q�_����G/2��N��;��;w��<{L�`�x�qJ�8�q]���J��nGF�{W�
��g�����!�������|!�w�v��T� ��O��᫂�����`8���k�˿S���s'�G�/	�H�?A�O��ǛX.v�z�H>5xS�����=é|R��<7o7�?�5H�,f����3����y[>f�9�|�/��ٲd*��U�����ۯj�hS�	NMI-������,e_o��V;JM���͏#I�s
6��`�A!OQ�
}�
��g�����!�����V?�?�����U�ܹ�����hrY<�.��Yc*�|���+(�[�,F8�O��}�
��g�����!����~��EUO�?^T����W}<c�婫�i��,��Mg�O/c�����g�pA,�/��6e�<�2@�"�L�7�FGFGRC+GPE~��W��������ψ4�����[_	�g�s2D���F�C�9��h|a�j���
�Y$��y���3rT��;T�ӌW�^U�7좥94���h�5�;|���5̮����G��<.z�Z�v��ͱ�e�_�}��f�vf����rO� ���*������E���C��UuJR�擻��k~ZzN�JN�G�i~2b��O"4�"������G�[�s	���+1�ϖO����s�
��g�����!����[x�n�T��v�[���������JMFpq��K>�ֵ�M���X$��ھ��O���$q�g�	
Y��\��֊�?��V>����?��'ԼU�Jg���߰ȷe�܁7�F݇vz�⊈�Ar��~�����T���|�����x�H�6p�z�kv�46'>[/�2��20~oP+�ʿL?���P��.��F�_��G�T���r�(��݅Xx��0��wI;0��q�zG�)�l��Ri��;����g*��֫���ܙ{8�O��?���U�O�x�����P���?Ƀ~ʟ����8-|�T`S��
{'��p}�5�h�Q綝�~��ۏZ6`d�”6�py�ܓM�8����u�(��e��5/�'��+����5>����=|�X=FT�ֲ]H5,kB[{
��7I����Wr�涆�)�m�vf�/'�R}�R������ 
��#P��UFvs�94�Ķ�$��D�~������3x�)�����"����S��e�w����EgEf�0<C�uK��pE,��?�tu����
�.��i���&���@o"���6� ��fU��`1���^�h-U7��u)c��ې���;�rn\���R�tV!Ǜ欵����~�.u��
�O���њ3Z�*;}ZӬ�6Q�3Tb}_���~ʘ�H�O���TX�2s�Q_X|uE����C8O����q�A�������3}�]�E��½*8f�:��W����%�lBd��:�tf���J��;X(�4f�����i?�����c�	���¾b���e�yq���Z��{+��'/Ø�
N=*!�mM�yp-�ی�ğ¾�ᮛ�5���2Kkv]ґ���?{�I�c�s�'��FV���v;�ƚ�<�3���5D�3�����1��NI�C���0`2:L�K���kN����E�5&��S�M����7�����Edd��|[�Ys�����Gج���

�^�N��ɣ|N��O�Ȗr��Jo��3�+�y�=y�˿�x'�����
,?��}S��������w��6:�Ϡ������~?�(�������?.C��;-���Jl?��|�[:±�5�f��O-�g��f���O����.�7��M���{Q���=�rU<zUq��"���W�9��rz�n�j���R��>�:�F�(�8$��G�o�����օ�~��*��׼C7�WS��δo�/�8�F���BG�{���~���"S�#;�Əa����}k����7��_)�ӂ���¼#M��M�w��c��}����C���E���V�s�n�eռ1����s0��{i�a�#���W�U�
-�_Y�TK�؃��m���ƫ�I8�;�7�֕*pm7%�k�c��lo�1[����A#?����5�.���8�Щ��ǔ�@�JlO>�W7~-k1��7�\��g�4a�Z�R������U�����l���߶���C�>3���?��������:���$;�6��A��������~?�(����x��Q������K�I��
�O�>����|}��������W�����1��J��T�TW��&�f���_ξ�������f J���9��׏�U�+�egty��GN��Ko�����[�Qm��dRrG�K-?��y�_ۏ�^���>��@8Ҭ�9�ҝu�lX�8�ƾ~��&d�K�?:�r���8���]���������J���0���ĺ׍u��jv�&��)��a��lH����$g��(�3��3�2�������"��������-���?�����ѱ��}~��o���.��U�����3���Nd*�R˜��2@����vF�r��Ow)Oڶ^�P������?�;����N�dG�j��������#�N4�C�����`߲��|x��N_(W������eO���O���P����'����ٺM���_.��Rg�95�>�����h�f0������[�q�ƾc��OZ���rj��w�~��qԹ)������דh���"���7"[Ȑ����W�kѻ
s���J��W�ɜ��}��Q[d�2L?+�����?�O�_�>j_�O_(W��8R��M_�X��w�t������k���+�o��_���c�FV�Cw,͖$�-��W��%��5kknv���죓�WҚ
i9��a^k6�-���<|l��Ÿ����{T��6��1W������ 99,����>4��0�
�W����+������W����[$w��q_3�K���&�����?�Ί?����l�.��f���"���ä�n��w����<-��^(�4�l'��-F���D�#��1���W��E����/�?���X4��g�?�~��G~&|d��{���@�ԃ�r�R^��Z�{x�ض1��>7ڱ��=;�j"�5�:c��~�K��~C��H�n�J�H��?�ւ��h��/���k����PR�tq5hiM������(x����:
����j��9���R��
��k��zU$���jT�YsM�~zx��	����	~G��	��uo�\��Na�w�p�H#n��=rk��	�v�f_x5�
�tI@P0N9���N1�~�zP+ѧ7y#�xzu4���k?�#��#�&�%�dž�X����,H>b��<��>����oũ�<y���I�ƿL����n�~'L�Y�q�Y�Y����g����"hq��������"����^���3��~����F�hB�yi�#���'���ٛ�
4���V�w�K�Oe+�$S"����`T�+�O�%�Ăo$��^��vWF�ǘ,l
�;s�B�׎�?Mj;
¦�YsN)��5%d��?���W��
y7�=yHږ2�\�q�w��,|S��7��?��Z���*j�iV|���q<=9M�kV~^���	�-h7��D����9���&?�"?�<�e���:wg0'���W���:P��m(ꏜao�[Q���k^Ե�mr[�jMU'�FUEh �a�9���Q_Gb�Ա}h�Œ{P
Q]Lm���F�Կ�6p3�MK��uo��ZO���
�����ZIx�1��1�.�䝧R(��*����Kأ��u��xG�;<֤7v|���県���RԵ/
ܝWR�W���5;!yqI$��{4Q�"&B"���O95�~��
о���k۫��n
�%`��d�lD�q���ȳ��ǭ����;�t���~���;\[���m����s���yU�2�0�I��u���U��hړi�x���'k}�8�����F}�?SHꚵ�B�cڏN)��:P(ǰ��(ǵ���ڌ{Q�jcڊL{R�ړ����5�Ҵ�n5+;�m"콳H�<�9f���:�ắ���)��ڻ�67-mqiy�N�٣��
.wd`��͇�|�$����kQ��7�γ^y�IJ�žS!�� �s�=O��:-+�z��-<�C�i�n$�B?��~��33v$�i
c��m:�K��l-ᵶ���#UP��
=Mh|��kV��f=o�NW4
�cڗԞ�U}F��:���Ț�ɍ��m�|�������
''dX���������G���I|�nP��!��I	�@Uwn`�
9��^�
CP�����-���-���-nB�n�9
�疾�>���׷�˘��[�%s��e���V1��۱�:�=��{�|�U"ܢ�j��Z���o��;�iq�^u��[F?h������a�TK11��Y�@�y`�rzע~�%ήyx�>G���a�j1�G�Gn�g(cڌ{P���ǵ&=��(�(ǵ~P
�G��E���^�^2��$w��Yi����kJ��p0��eF�Fx��
��7�CN���t�&��&zf��H�4^S:J��1��Cg#�]/��2�G�e����v��x�ZH�v�/���QEvz֟q�Z$vڭޒ����i;�����~x��kZ𮶺=�[x�[��B�8�P�6Ӏ]m(�hGj(���Ό:N��3�i:�׺v�����<�Fic`
��)�A�~1�j;L���O,�q��Ӥ�--�iwL�3��A�O,x���"��#��%���J:tH�m�kZ~�kga���F9ID;_iP<�p1�#�J��h� ���uw��^��!�H~�/�UH,M��䑁����ޔ���}70��XUUE)i��a���鶾Uƥu�ɸ��v�+��".?�֮� QEw�&��E�c$}(��t��(��
endstream
endobj
166 0 obj
<< /Type /Annot
/Subtype /Link
/A 167 0 R
/Border [0 0 0]
/H /I
/Rect [ 244.6402 625.0268 260.9032 634.9474 ]
>>
endobj
167 0 obj
<< /Type /Action
>>
endobj
168 0 obj
<< /Type /Annot
/Subtype /Link
/A 169 0 R
/Border [0 0 0]
/H /I
/Rect [ 127.5720 526.6936 146.5455 536.6142 ]
>>
endobj
169 0 obj
<< /Type /Action
>>
endobj
170 0 obj
<< /Type /Annot
/Subtype /Link
/A 171 0 R
/Border [0 0 0]
/H /I
/Rect [ 68.5162 490.9793 87.4898 500.8999 ]
>>
endobj
171 0 obj
<< /Type /Action
>>
endobj
172 0 obj
<< /Type /Annot
/Subtype /Link
/A 173 0 R
/Border [0 0 0]
/H /I
/Rect [ 363.8340 313.5893 399.0705 323.5100 ]
>>
endobj
173 0 obj
<< /Type /Action
>>
endobj
174 0 obj
<< /Type /Annot
/Subtype /Link
/A 175 0 R
/Border [0 0 0]
/H /I
/Rect [ 503.3857 246.5656 522.3592 256.4862 ]
>>
endobj
175 0 obj
<< /Type /Action
>>
endobj
176 0 obj
<< /Type /Annot
/Subtype /Link
/A 177 0 R
/Border [0 0 0]
/H /I
/Rect [ 86.9340 222.7561 105.9075 232.6767 ]
>>
endobj
177 0 obj
<< /Type /Action
>>
endobj
178 0 obj
<< /Type /Annot
/Subtype /Link
/A 179 0 R
/Border [0 0 0]
/H /I
/Rect [ 330.2647 222.7561 349.2382 232.6767 ]
>>
endobj
179 0 obj
<< /Type /Action
>>
endobj
180 0 obj
<< /Type /Annot
/Subtype /Link
/A 181 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 79.6223 147.7500 192.1223 ]
>>
endobj
181 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-1-new.jpg)
>>
endobj
182 0 obj
<< /Type /Annot
/Subtype /Link
/A 183 0 R
/Border [0 0 0]
/H /I
/Rect [ 244.6402 625.0268 260.9032 634.9474 ]
>>
endobj
183 0 obj
<< /Type /Action
>>
endobj
184 0 obj
<< /Type /Annot
/Subtype /Link
/A 185 0 R
/Border [0 0 0]
/H /I
/Rect [ 127.5720 526.6936 146.5455 536.6142 ]
>>
endobj
185 0 obj
<< /Type /Action
>>
endobj
186 0 obj
<< /Type /Annot
/Subtype /Link
/A 187 0 R
/Border [0 0 0]
/H /I
/Rect [ 68.5162 490.9793 87.4898 500.8999 ]
>>
endobj
187 0 obj
<< /Type /Action
>>
endobj
188 0 obj
<< /Type /Annot
/Subtype /Link
/A 189 0 R
/Border [0 0 0]
/H /I
/Rect [ 363.8340 313.5893 399.0705 323.5100 ]
>>
endobj
189 0 obj
<< /Type /Action
>>
endobj
190 0 obj
<< /Type /Annot
/Subtype /Link
/A 191 0 R
/Border [0 0 0]
/H /I
/Rect [ 503.3857 246.5656 522.3592 256.4862 ]
>>
endobj
191 0 obj
<< /Type /Action
>>
endobj
192 0 obj
<< /Type /Annot
/Subtype /Link
/A 193 0 R
/Border [0 0 0]
/H /I
/Rect [ 86.9340 222.7561 105.9075 232.6767 ]
>>
endobj
193 0 obj
<< /Type /Action
>>
endobj
194 0 obj
<< /Type /Annot
/Subtype /Link
/A 195 0 R
/Border [0 0 0]
/H /I
/Rect [ 330.2647 222.7561 349.2382 232.6767 ]
>>
endobj
195 0 obj
<< /Type /Action
>>
endobj
196 0 obj
<< /Type /Annot
/Subtype /Link
/A 197 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 79.6223 147.7500 192.1223 ]
>>
endobj
197 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-1-new.jpg)
>>
endobj
198 0 obj
<< /Type /Page
/Parent 3 0 R
/Annots [ 200 0 R 202 0 R 204 0 R 206 0 R 209 0 R 212 0 R 214 0 R 216 0 R 218 0 R 220 0 R 222 0 R 224 0 R 226 0 R 228 0 R 230 0 R ]
/Contents 199 0 R
>>
endobj
199 0 obj
<<
/Length 18214 >>
stream

q
15.000 33.143 577.500 743.857 re W n
0.965 0.965 0.965 rg
26.250 671.664 555.000 105.336 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 671.664 m 581.250 671.664 l 581.250 672.414 l 26.250 672.414 l  f
q
35.250 682.914 537.000 94.086 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 1: Identification and quantification of human satellite cells on single fibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(All the nuclei of fragments of human myofibres of a parispinal muscle biopsy of a healthy adolescent patient were )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(recognized by the human specific lamin A/C antibody \(A I\). Along the fibres, several nuclei were detected by Pax7 antibody )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(\(A II\) and were in the classical satellite cell position beneath the basal lamina \(A III\). Nuclei were counterstained with DAPI )] TJ ET
BT 35.250 706.897 Td /F1 9.8 Tf  [(and merged pictures are shown in panels \(A I, II, III\). \(B\) Number of satellite cells per 100�m fibre length was calculated. )] TJ ET
BT 35.250 693.161 Td /F1 9.8 Tf  [(Scale bars: 10�m.)] TJ ET
Q
BT 26.250 654.640 Td /F4 9.8 Tf  [(Human and mouse satellite cells are defined by a different marker signature)] TJ ET
BT 26.250 635.235 Td /F1 9.8 Tf  [(As Pax7 characterizes the majority of satellite cells in mouse )] TJ ET
0.267 0.267 0.267 rg
BT 290.134 635.235 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 300.976 635.235 Td /F1 9.8 Tf  [(, and appears to be frequently expressed on human fibres )] TJ ET
BT 26.250 623.331 Td /F1 9.8 Tf  [(\(Figure 1A\), we wished to determine if human satellite cells are identified by antibodies to other proteins expressed by mouse )] TJ ET
BT 26.250 611.426 Td /F1 9.8 Tf  [(satellite cells. Caveolin-1, a surface protein expressed by virtually all mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 413.130 611.426 Td /F1 9.8 Tf  [([13] )] TJ ET
0.271 0.267 0.267 rg
BT 432.103 611.426 Td /F1 9.8 Tf  [(\(Figure 2A\), was also expressed )] TJ ET
BT 26.250 599.521 Td /F1 9.8 Tf  [(on a sub-population of human satellite cells \(Figure 2B\). M-cadherin was expressed on the cell membrane of both mouse and )] TJ ET
BT 26.250 587.616 Td /F1 9.8 Tf  [(human satellite cells \(Figure 2B\). We confirmed that CD34, that was reported to be expressed by mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 536.156 587.616 Td /F1 9.8 Tf  [([11])] TJ ET
0.271 0.267 0.267 rg
BT 552.418 587.616 Td /F1 9.8 Tf  [(, was )] TJ ET
BT 26.250 575.712 Td /F1 9.8 Tf  [(indeed expressed on mouse satellite cells under the basal lamina \(Figure 2E\), but this antibody did not stain human satellite )] TJ ET
BT 26.250 563.807 Td /F1 9.8 Tf  [(cells, possibly because of differences between the murine and human epitopes. Conversely, the human CD29 antibody )] TJ ET
BT 26.250 551.902 Td /F1 9.8 Tf  [(recognised human, but not mouse satellite cells \(Figure 2F\).)] TJ ET
0.965 0.965 0.965 rg
26.250 267.226 555.000 274.795 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 542.021 m 581.250 542.021 l 581.250 541.271 l 26.250 541.271 l  f
26.250 267.226 m 581.250 267.226 l 581.250 267.976 l 26.250 267.976 l  f
q
112.500 0 0 112.500 35.250 419.771 cm /I4 Do
Q 
q
35.250 278.476 537.000 135.295 re W n
0.271 0.267 0.267 rg
BT 35.250 404.247 Td /F4 9.8 Tf  [(Fig. 2: Characterization of mouse and human satellite cells on single fibres.)] TJ ET
BT 35.250 384.877 Td /F1 9.8 Tf  [(Detection of mouse \(arrows\) and human \(arrowheads\) satellite cells on single fibres. Myofibre fragments of adult 3F-nLacZ-)] TJ ET
BT 35.250 371.141 Td /F1 9.8 Tf  [(2E mouse tibialis anterior muscle were used in order to clearly distinguish DAPI positive satellite cells from X-Gal positive )] TJ ET
BT 35.250 357.405 Td /F1 9.8 Tf  [(myonuclei. Pax7 antibody recognizes the majority of satellite cells on both mouse \(A\) and human \(B\) fibre fragments. )] TJ ET
BT 35.250 343.669 Td /F1 9.8 Tf  [(Caveolin-1 is expressed both in mouse \(A\) and human \(B\) satellite cells. M-Cadherin is expressed in the membrane of both )] TJ ET
BT 35.250 329.932 Td /F1 9.8 Tf  [(mouse \(C\) and human \(D\) satellite cells. The antibody raised to mouse CD34 indeed labels only mouse satellite cells \(E\), )] TJ ET
BT 35.250 316.196 Td /F1 9.8 Tf  [(whilst CD29 is a human specific antibody that reacts only with human satellite cells \(F\). Laminin immunostaining was )] TJ ET
BT 35.250 302.460 Td /F1 9.8 Tf  [(performed in combination with satellite cell markers wherever possible and DAPI was used for counterstaining mouse )] TJ ET
BT 35.250 288.724 Td /F1 9.8 Tf  [(satellite cells and human nuclei. Scale bars: 10�m.)] TJ ET
Q
BT 26.250 250.203 Td /F4 9.8 Tf  [(Fragility of DMD myofibres does not allow dissection of single fibres)] TJ ET
BT 26.250 230.798 Td /F1 9.8 Tf  [(We were not able to prepare fibre fragments derived from DMD muscles using the method of myofibre isolation that we )] TJ ET
BT 26.250 218.893 Td /F1 9.8 Tf  [(successfully applied to normal human muscle \(Figure 3A\). The fragility of fibres from DMD muscle caused them to hyper-)] TJ ET
BT 26.250 206.988 Td /F1 9.8 Tf  [(contract before the isolation and to break during the isolation process. Furthermore, the interference of connective tissue )] TJ ET
BT 26.250 195.084 Td /F1 9.8 Tf  [(prevented single fibre isolation and only rarely could bundles of fibres be isolated for immunostaining \(Figure 3B and 3C\). )] TJ ET
BT 26.250 183.179 Td /F1 9.8 Tf  [(Although our technique did not allow a clear identification of human satellite cells, Pax7 is expressed by a few nuclei in this )] TJ ET
BT 26.250 171.274 Td /F1 9.8 Tf  [(preparation of DMD muscle \(Figure 3B and C\).)] TJ ET
0.965 0.965 0.965 rg
26.250 33.143 555.000 128.250 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 161.393 m 581.250 161.393 l 581.250 160.643 l 26.250 160.643 l  f
q
112.500 0 0 112.500 35.250 39.143 cm /I5 Do
Q 
q
35.250 33.143 537.000 0.000 re W n
Q
Q
q
15.000 33.143 577.500 743.857 re W n
0.965 0.965 0.965 rg
26.250 671.664 555.000 105.336 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 671.664 m 581.250 671.664 l 581.250 672.414 l 26.250 672.414 l  f
q
35.250 682.914 537.000 94.086 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 1: Identification and quantification of human satellite cells on single fibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(All the nuclei of fragments of human myofibres of a parispinal muscle biopsy of a healthy adolescent patient were )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(recognized by the human specific lamin A/C antibody \(A I\). Along the fibres, several nuclei were detected by Pax7 antibody )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(\(A II\) and were in the classical satellite cell position beneath the basal lamina \(A III\). Nuclei were counterstained with DAPI )] TJ ET
BT 35.250 706.897 Td /F1 9.8 Tf  [(and merged pictures are shown in panels \(A I, II, III\). \(B\) Number of satellite cells per 100�m fibre length was calculated. )] TJ ET
BT 35.250 693.161 Td /F1 9.8 Tf  [(Scale bars: 10�m.)] TJ ET
Q
BT 26.250 654.640 Td /F4 9.8 Tf  [(Human and mouse satellite cells are defined by a different marker signature)] TJ ET
BT 26.250 635.235 Td /F1 9.8 Tf  [(As Pax7 characterizes the majority of satellite cells in mouse )] TJ ET
0.267 0.267 0.267 rg
BT 290.134 635.235 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 300.976 635.235 Td /F1 9.8 Tf  [(, and appears to be frequently expressed on human fibres )] TJ ET
BT 26.250 623.331 Td /F1 9.8 Tf  [(\(Figure 1A\), we wished to determine if human satellite cells are identified by antibodies to other proteins expressed by mouse )] TJ ET
BT 26.250 611.426 Td /F1 9.8 Tf  [(satellite cells. Caveolin-1, a surface protein expressed by virtually all mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 413.130 611.426 Td /F1 9.8 Tf  [([13] )] TJ ET
0.271 0.267 0.267 rg
BT 432.103 611.426 Td /F1 9.8 Tf  [(\(Figure 2A\), was also expressed )] TJ ET
BT 26.250 599.521 Td /F1 9.8 Tf  [(on a sub-population of human satellite cells \(Figure 2B\). M-cadherin was expressed on the cell membrane of both mouse and )] TJ ET
BT 26.250 587.616 Td /F1 9.8 Tf  [(human satellite cells \(Figure 2B\). We confirmed that CD34, that was reported to be expressed by mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 536.156 587.616 Td /F1 9.8 Tf  [([11])] TJ ET
0.271 0.267 0.267 rg
BT 552.418 587.616 Td /F1 9.8 Tf  [(, was )] TJ ET
BT 26.250 575.712 Td /F1 9.8 Tf  [(indeed expressed on mouse satellite cells under the basal lamina \(Figure 2E\), but this antibody did not stain human satellite )] TJ ET
BT 26.250 563.807 Td /F1 9.8 Tf  [(cells, possibly because of differences between the murine and human epitopes. Conversely, the human CD29 antibody )] TJ ET
BT 26.250 551.902 Td /F1 9.8 Tf  [(recognised human, but not mouse satellite cells \(Figure 2F\).)] TJ ET
0.965 0.965 0.965 rg
26.250 267.226 555.000 274.795 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 542.021 m 581.250 542.021 l 581.250 541.271 l 26.250 541.271 l  f
26.250 267.226 m 581.250 267.226 l 581.250 267.976 l 26.250 267.976 l  f
q
112.500 0 0 112.500 35.250 419.771 cm /I4 Do
Q 
q
35.250 278.476 537.000 135.295 re W n
0.271 0.267 0.267 rg
BT 35.250 404.247 Td /F4 9.8 Tf  [(Fig. 2: Characterization of mouse and human satellite cells on single fibres.)] TJ ET
BT 35.250 384.877 Td /F1 9.8 Tf  [(Detection of mouse \(arrows\) and human \(arrowheads\) satellite cells on single fibres. Myofibre fragments of adult 3F-nLacZ-)] TJ ET
BT 35.250 371.141 Td /F1 9.8 Tf  [(2E mouse tibialis anterior muscle were used in order to clearly distinguish DAPI positive satellite cells from X-Gal positive )] TJ ET
BT 35.250 357.405 Td /F1 9.8 Tf  [(myonuclei. Pax7 antibody recognizes the majority of satellite cells on both mouse \(A\) and human \(B\) fibre fragments. )] TJ ET
BT 35.250 343.669 Td /F1 9.8 Tf  [(Caveolin-1 is expressed both in mouse \(A\) and human \(B\) satellite cells. M-Cadherin is expressed in the membrane of both )] TJ ET
BT 35.250 329.932 Td /F1 9.8 Tf  [(mouse \(C\) and human \(D\) satellite cells. The antibody raised to mouse CD34 indeed labels only mouse satellite cells \(E\), )] TJ ET
BT 35.250 316.196 Td /F1 9.8 Tf  [(whilst CD29 is a human specific antibody that reacts only with human satellite cells \(F\). Laminin immunostaining was )] TJ ET
BT 35.250 302.460 Td /F1 9.8 Tf  [(performed in combination with satellite cell markers wherever possible and DAPI was used for counterstaining mouse )] TJ ET
BT 35.250 288.724 Td /F1 9.8 Tf  [(satellite cells and human nuclei. Scale bars: 10�m.)] TJ ET
Q
BT 26.250 250.203 Td /F4 9.8 Tf  [(Fragility of DMD myofibres does not allow dissection of single fibres)] TJ ET
BT 26.250 230.798 Td /F1 9.8 Tf  [(We were not able to prepare fibre fragments derived from DMD muscles using the method of myofibre isolation that we )] TJ ET
BT 26.250 218.893 Td /F1 9.8 Tf  [(successfully applied to normal human muscle \(Figure 3A\). The fragility of fibres from DMD muscle caused them to hyper-)] TJ ET
BT 26.250 206.988 Td /F1 9.8 Tf  [(contract before the isolation and to break during the isolation process. Furthermore, the interference of connective tissue )] TJ ET
BT 26.250 195.084 Td /F1 9.8 Tf  [(prevented single fibre isolation and only rarely could bundles of fibres be isolated for immunostaining \(Figure 3B and 3C\). )] TJ ET
BT 26.250 183.179 Td /F1 9.8 Tf  [(Although our technique did not allow a clear identification of human satellite cells, Pax7 is expressed by a few nuclei in this )] TJ ET
BT 26.250 171.274 Td /F1 9.8 Tf  [(preparation of DMD muscle \(Figure 3B and C\).)] TJ ET
0.965 0.965 0.965 rg
26.250 33.143 555.000 128.250 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 161.393 m 581.250 161.393 l 581.250 160.643 l 26.250 160.643 l  f
q
112.500 0 0 112.500 35.250 39.143 cm /I5 Do
Q 
q
35.250 33.143 537.000 0.000 re W n
Q
Q
q
15.000 33.143 577.500 743.857 re W n
0.965 0.965 0.965 rg
26.250 671.664 555.000 105.336 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 671.664 m 581.250 671.664 l 581.250 672.414 l 26.250 672.414 l  f
q
35.250 682.914 537.000 94.086 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 1: Identification and quantification of human satellite cells on single fibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(All the nuclei of fragments of human myofibres of a parispinal muscle biopsy of a healthy adolescent patient were )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(recognized by the human specific lamin A/C antibody \(A I\). Along the fibres, several nuclei were detected by Pax7 antibody )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(\(A II\) and were in the classical satellite cell position beneath the basal lamina \(A III\). Nuclei were counterstained with DAPI )] TJ ET
BT 35.250 706.897 Td /F1 9.8 Tf  [(and merged pictures are shown in panels \(A I, II, III\). \(B\) Number of satellite cells per 100�m fibre length was calculated. )] TJ ET
BT 35.250 693.161 Td /F1 9.8 Tf  [(Scale bars: 10�m.)] TJ ET
Q
BT 26.250 654.640 Td /F4 9.8 Tf  [(Human and mouse satellite cells are defined by a different marker signature)] TJ ET
BT 26.250 635.235 Td /F1 9.8 Tf  [(As Pax7 characterizes the majority of satellite cells in mouse )] TJ ET
0.267 0.267 0.267 rg
BT 290.134 635.235 Td /F1 9.8 Tf  [([9])] TJ ET
0.271 0.267 0.267 rg
BT 300.976 635.235 Td /F1 9.8 Tf  [(, and appears to be frequently expressed on human fibres )] TJ ET
BT 26.250 623.331 Td /F1 9.8 Tf  [(\(Figure 1A\), we wished to determine if human satellite cells are identified by antibodies to other proteins expressed by mouse )] TJ ET
BT 26.250 611.426 Td /F1 9.8 Tf  [(satellite cells. Caveolin-1, a surface protein expressed by virtually all mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 413.130 611.426 Td /F1 9.8 Tf  [([13] )] TJ ET
0.271 0.267 0.267 rg
BT 432.103 611.426 Td /F1 9.8 Tf  [(\(Figure 2A\), was also expressed )] TJ ET
BT 26.250 599.521 Td /F1 9.8 Tf  [(on a sub-population of human satellite cells \(Figure 2B\). M-cadherin was expressed on the cell membrane of both mouse and )] TJ ET
BT 26.250 587.616 Td /F1 9.8 Tf  [(human satellite cells \(Figure 2B\). We confirmed that CD34, that was reported to be expressed by mouse satellite cells )] TJ ET
0.267 0.267 0.267 rg
BT 536.156 587.616 Td /F1 9.8 Tf  [([11])] TJ ET
0.271 0.267 0.267 rg
BT 552.418 587.616 Td /F1 9.8 Tf  [(, was )] TJ ET
BT 26.250 575.712 Td /F1 9.8 Tf  [(indeed expressed on mouse satellite cells under the basal lamina \(Figure 2E\), but this antibody did not stain human satellite )] TJ ET
BT 26.250 563.807 Td /F1 9.8 Tf  [(cells, possibly because of differences between the murine and human epitopes. Conversely, the human CD29 antibody )] TJ ET
BT 26.250 551.902 Td /F1 9.8 Tf  [(recognised human, but not mouse satellite cells \(Figure 2F\).)] TJ ET
0.965 0.965 0.965 rg
26.250 267.226 555.000 274.795 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 542.021 m 581.250 542.021 l 581.250 541.271 l 26.250 541.271 l  f
26.250 267.226 m 581.250 267.226 l 581.250 267.976 l 26.250 267.976 l  f
q
112.500 0 0 112.500 35.250 419.771 cm /I4 Do
Q 
q
35.250 278.476 537.000 135.295 re W n
0.271 0.267 0.267 rg
BT 35.250 404.247 Td /F4 9.8 Tf  [(Fig. 2: Characterization of mouse and human satellite cells on single fibres.)] TJ ET
BT 35.250 384.877 Td /F1 9.8 Tf  [(Detection of mouse \(arrows\) and human \(arrowheads\) satellite cells on single fibres. Myofibre fragments of adult 3F-nLacZ-)] TJ ET
BT 35.250 371.141 Td /F1 9.8 Tf  [(2E mouse tibialis anterior muscle were used in order to clearly distinguish DAPI positive satellite cells from X-Gal positive )] TJ ET
BT 35.250 357.405 Td /F1 9.8 Tf  [(myonuclei. Pax7 antibody recognizes the majority of satellite cells on both mouse \(A\) and human \(B\) fibre fragments. )] TJ ET
BT 35.250 343.669 Td /F1 9.8 Tf  [(Caveolin-1 is expressed both in mouse \(A\) and human \(B\) satellite cells. M-Cadherin is expressed in the membrane of both )] TJ ET
BT 35.250 329.932 Td /F1 9.8 Tf  [(mouse \(C\) and human \(D\) satellite cells. The antibody raised to mouse CD34 indeed labels only mouse satellite cells \(E\), )] TJ ET
BT 35.250 316.196 Td /F1 9.8 Tf  [(whilst CD29 is a human specific antibody that reacts only with human satellite cells \(F\). Laminin immunostaining was )] TJ ET
BT 35.250 302.460 Td /F1 9.8 Tf  [(performed in combination with satellite cell markers wherever possible and DAPI was used for counterstaining mouse )] TJ ET
BT 35.250 288.724 Td /F1 9.8 Tf  [(satellite cells and human nuclei. Scale bars: 10�m.)] TJ ET
Q
BT 26.250 250.203 Td /F4 9.8 Tf  [(Fragility of DMD myofibres does not allow dissection of single fibres)] TJ ET
BT 26.250 230.798 Td /F1 9.8 Tf  [(We were not able to prepare fibre fragments derived from DMD muscles using the method of myofibre isolation that we )] TJ ET
BT 26.250 218.893 Td /F1 9.8 Tf  [(successfully applied to normal human muscle \(Figure 3A\). The fragility of fibres from DMD muscle caused them to hyper-)] TJ ET
BT 26.250 206.988 Td /F1 9.8 Tf  [(contract before the isolation and to break during the isolation process. Furthermore, the interference of connective tissue )] TJ ET
BT 26.250 195.084 Td /F1 9.8 Tf  [(prevented single fibre isolation and only rarely could bundles of fibres be isolated for immunostaining \(Figure 3B and 3C\). )] TJ ET
BT 26.250 183.179 Td /F1 9.8 Tf  [(Although our technique did not allow a clear identification of human satellite cells, Pax7 is expressed by a few nuclei in this )] TJ ET
BT 26.250 171.274 Td /F1 9.8 Tf  [(preparation of DMD muscle \(Figure 3B and C\).)] TJ ET
0.965 0.965 0.965 rg
26.250 33.143 555.000 128.250 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 161.393 m 581.250 161.393 l 581.250 160.643 l 26.250 160.643 l  f
q
112.500 0 0 112.500 35.250 39.143 cm /I5 Do
Q 
q
35.250 33.143 537.000 0.000 re W n
Q
Q
q
112.500 0 0 112.500 35.250 419.771 cm /I4 Do
Q 
q
112.500 0 0 112.500 35.250 39.143 cm /I5 Do
Q 
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(3)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
200 0 obj
<< /Type /Annot
/Subtype /Link
/A 201 0 R
/Border [0 0 0]
/H /I
/Rect [ 290.1338 634.3336 300.9758 644.2542 ]
>>
endobj
201 0 obj
<< /Type /Action
>>
endobj
202 0 obj
<< /Type /Annot
/Subtype /Link
/A 203 0 R
/Border [0 0 0]
/H /I
/Rect [ 413.1300 610.5241 432.1035 620.4447 ]
>>
endobj
203 0 obj
<< /Type /Action
>>
endobj
204 0 obj
<< /Type /Annot
/Subtype /Link
/A 205 0 R
/Border [0 0 0]
/H /I
/Rect [ 536.1555 586.7146 552.4185 596.6352 ]
>>
endobj
205 0 obj
<< /Type /Action
>>
endobj
206 0 obj
<< /Type /Annot
/Subtype /Link
/A 207 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 419.7712 147.7500 532.2712 ]
>>
endobj
207 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-2-new.jpg)
>>
endobj
208 0 obj
<<
/Type /XObject
/Subtype /Image
/Width 150
/Height 150
/ColorSpace /DeviceRGB
/Filter /DCTDecode
/BitsPerComponent 8
/Length 9833>>
stream
����JFIF



��;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90
��C




��C
		

����
"

��







	
���

}
!1AQa"q2���#B��R��$3br�	
%&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz���������������������������������������������������������������������������










	
���

w
!1AQaq"2�B����	#3R�br�
$4�%�&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz��������������������������������������������������������������������������
?����.�g��^$]���ZC>ff�`��
t|�F9̒u���֗aoq��{p��(-!n���>����u�G�|����[9I�ʟ���;|���5�V��8�\�n�nt��g�'��NW#zHX�:��M����挧��J�[S��^�y{�׈�Y���g��u�6Z���8t��ԩ/<�;J��(S��Eji>��m扫�Ga�n�¬�z����^�g���]�:�����u)�'7��̲ٻʩ�0	�UB���J���)JKT��Z6��jc�����V /����}�+�G��|#uk��>��f�E�
>;UT�<�&C!;���_��2�����A�1JH������zԔ&+���E�Ğ	}"�mo��k+Dp�=�A ���q_˵S7���y����"�?������H�]���z��m���Ig�� 
����A�p�kj�S��'�'��,���ua��O�??15r�,�wC�*�!\dU�n��6�9�e�&�p#w�@�������u��s[U����������\����^��="��_����;x(`����޿��W���Ԧ�'	n���ڢV�x�z��ӂ+SU���%�k�uP��+\[��#�h��P���
��,�cWbȼVq���U��?F���W>#���(S�����>#za����{��8�3�����k��Z�v��V0mT�O�ҾO�<9��!{)v�d���O�
q���_�O��t�ۍ:�)4�6��R&upp�A�ƿ
�ş��2�	�����]%�|����������O�._�y��k��~L
и���������m���"�3Ω?�O��w�dp��:��s�ρ���7��SwV(��5�H�#���͹�
s�%��������X���˫i�0c��08�����G��a{�C�2��70�ġ���f�\yu�]����H��JuV ����q��U%ed˧^)�8�-�����曥���z���V��!�	mVit���7]�Wr|�T�FN<�G�!����/���+�����$k$��Wm����_��%\kZ�Ř.�^=�XqZ�c)&�>��7_�������P:>F�b@��E~9�x�ŲM"�_U�5I�?��
��_�y&�؞O����R%4�?�^�]��e�d��óQ%���/��P��9�t�"��y
ʍ+J�g�q��p��0���k��+G��O����t�j�]�Ь�ȳ���&N�`2k����rs�	��rOO�I�����8��^�J
O���t����Cu��
�_�	'�T���d��@g��$�
��g��9������t�����~ �����]iB���U)�MZOFw�ׅ5Y��&;�+Sھ'ם�;W.
��v�2A�����W�[�Aa�����O�J�NG\�<g�:{סk?<d��Wź��?�5��a�M��m4��N�1���\L�k"��2�����Nl�_J�o�؇Rr�o�6�EN��"~z�~6��0?�
�����$��OO��3Ƈ��$��]�<o2�3�Շ�E'���-�[�JN|e�3��?�O���~��e4�Я�"����~!=�uf��B� h�>x�dQR�<g��V�����Z�Qh��i}y$ʇ� 184WV.rswg�_�T��1��}?�i��q������+���
{d���S�X�Q�K�Y�u+a�2*�.
5��S�2��T%w����ϵg:���'c��jt����2㵸�TKdi��/QP���J�V�8�I���
h��w�D�_�$�M�Mn����`+9]�����c����i�2�2��SX����
���S��3q0FH?g��Hi=��,�,��1�r��kɿc�9�
��Կ�"J��M��0�ͻ���v08K(#py��ҝLE,*^���p����ҮR�%�K5€�����pMh��-��-u4�<���@
���䌏ā]
�����r%����X�;�`�7�Xs��g��ii����W�Ҝ5LN�Z36����I<�{2͙���/�)�O�_�V��m��Z�*����t��+����k��E.a����X/�<	?�
c�寉���g���$`�3��p95�����W�!��e������f�$b� H�'W��#88�+�Q�Z��ݬ�*�9�C���d3������2�JҒL�"�,��0��-�mF��p�{i+[�$R�ɽN��*� �	� �~:��ږ�.�������+���ۜ����סD�ֱi��h���=]��|�?�U�܏���+���4��x��W���n��(�[̽��fє�n�����Z����㵋Ck�R�J���$����$c,E(K�RI�(I��e�Xc��TEz�Er�L�*��@钤�m���
�p����2����J�i|`�#�����f�}�)^=�L��{��m���`����;~�QW����~*|@P�C������4W����!������ �bM'�z_��?e��_�n���
ǟ���5���,eX�������_��
[��²�Rt.zq����8��� �������yB@�����3������
E�����'G�^��_�&\�d�������^��aX��$� F�J�_�6?�n���W���<+
?ނν��d`թ|GeA�J������a����G���
=:?|��7�����@��ҭ./�{=�)��vꀂB��z�a?��c`W�3� �K
�ҽ#�儞�$Ku�}RH�7,E�Ā?_��{��Ȩ��c�T�X��%K[��+G����.q����
����,���  �G_j��;�5�{R�kv���場�ˆ"�[j/�����M=�K^��}���������|�i0��������k0xg�ږ�<RIm���u$P
��4,B�@� {��_�\k�i���.��
��n-����	�I��
I�]H�#�5n�m5�??�j�ۓ���/�gG��8|e��@Ϩ^��J�e�#i��~�G<W��)��F�4��i��Ex���������=�Q�C��V8a@��2p�
�����{׻��FpS��_�1r���5�Q�٧?�>�;��#L��(���~��4i�#?��?����i�-�*).��+����wn��NL�]�?�߳ŷ�}Z��v��K�յ84��8�f�7�\l���G'��+/�(���$f?�f�=c'qS��'�+���Y�golc�Z���i��!�|��v���Y)5}/U�M�~���
��$��4��'�������kd��5i�Ώ�����ڄP����S���D���\�u��=9�2�1�Ik�~�~���¯��1�&����e�������������(BaPN<�E|��L��������(Wr��>Z���tՑ�e�I
�
o]�c�[��g�|~�W�v�T�iM�p��Ka��9������4���>(�u�Et�6ݭD���#�u�Ȁ��03��������y�:�ro]�M�_����V�J����9e9E��;��-5i�X�,d��Bʅn���ߡ�:�V]9O���P4�nx/'�'��Uko�hw�H*#�o�8��sR��a�$�8>3�ﹳN�M�S��b��Qis{F�=gU��O��ʊH�E��n�X�me�,��	�_�*pFk��A����E���Z7���R�o������Ye6��q0ch$�8���,6��|.�#�P��g�C���f���'{�oe�<�v��D|�
��OUF'-�_C��ߥ�
|[�p�8��Ͽ�y�R���b��<>=�Ųc�j-�����)I�k�=N�5�H��K��<_��n�t�.c�ЅOݺW�ݢ��bb���bxi��m��NӵB#��^��Z��qU!Rܧ���
����
�@�%|DY�n�}+��
��xZ����Mk_��n��h>�oxϷ���A�7s�Ҿ9ռ�MV�K�X
��� �d���^��ϵzP�d�jU����:�2���{��sʄ
FY��G�	�W������>&�|��?��?�E���-�y~'x~2Fp�:�q��-k��Z�"�F�Eh������w�n��k���&�^m%�����M߂|+c�;�����C��_�n�=�����i9P�`T(�@8�88崿�>`��9�������.�t�Ϲ5�W�YJ�4��a���i��+���{�n������T0��������J1�BO=�k��u*�K��W��ZG������Ly�M͵
=HEcӤ�U�k�b�4��-��Ω������P_���!�����!8��C?�+X(Tj����Pj0��}}��)~��E{������n�!�Q]O������׵�v��9���W����`N>bf�
�$�Z�4�#����G�.�ӿ�T���|C��4޿�����?�?Z�����l��%����|�7��b��K���:��T�8>ՕI�=6�W�1E���'�G�'�d�k���)�֒8��dc�M�I�
�c`y$5��5����{3���"t��������?��j�ȷ:oXه�0Z�٦ٓ�o��)Uw����,�찼fdR��;�3��L�ʥ���O�\X�����I�g��Z�֥��}�ռ"���w�'�0�2Ī�8�
4Ț�YH�9���8Ϩ5 �+)�ye�=�$�����Kڮ�lG��5+7�i�0ݏ"�q�:�A����{r-�
�e���pG+�ç�|8������~+�Y�Ժ����"_�;!�'����u["@Spޤ�~��������Erſ�l�{�_}�k�
�Dx$�z�)�N�CȩFs����:s��cB�
��5�ǭc_�h���y���9?��L���..^@9؇��+hNB��w�A���cJ����JZ���i����煞Y��X�U������x�Q�L�4�f�|t*�_�A����
����F��
��zi5��^,��u��
��3�5��p�MCB�(�RG���T��ʁ"�_TQ��s�`k~8����yw�'�c�}�"
$kz
�7~e���9�t��"�1�7��@�7��/iy���ZȰ����"A����^@���
���h�:���+^ެW��k��}��]ե�~1|Gg�Ph��_O�Ej���'���O�2�L�&�
���ph��m�{��7����?�f_x3��?�:֥5��o��V;�$�Dc��8�i���b���17��a�����Tp�5���3^�|7��b=A�#���ٷ�1…�V�f$�������P��~,ZY�WR��P�}��
ۗ<����ǧ>�C,S�f�����J��w�k���υu�&�mv����[���֜�6O�G�9����2�i}�[��:��N����h�E�x�cYgo�[�NN1��r9<���I��㯵]�B�	&�"�����v�"0NJ�������\��Bm6��ȈӦյ>�)������3H��_%���\	$�ٱ|��af'�`8�����:M>6]AtʞdcQ�`�h߆�
�v'q�����|i��Cė��Ǘsi���B�e��;y0p���w�=�������-7���̭�z%6�-����������>���b�����(ז�)K��!,?�2�����nY>
���j�~�uy4�P���D��_�;% H�4]=�}����w����ӧ�{O�om��0ɡ�Ŕ��޸p���j�m��ߗ���dž��n�u�yZOFޏ���?��Ѻg�C��_	���I+�Y��=�7#����������=���{(e���)'Du���FA`��z���T:�υ�x�K��֤��lgu��P�؂ 6���*;"��Ϳ�('�%�;������~�
މ���������$�V9'���_;E����me,QxJ0��u=?�9��OA�����P��?�����/�CǍc�G�Y�8:E��l��{�H\��.p+�&����Ӄ8��
�F����5\�Uf������+�s���~���c�����D�o�8�g]���H4���A���c޴������R
7��7������������z�7�������M��jO�z'�5�E+�(Zo�#V3�	|H���Կ2=�_�&��_�7𦅬x"���u�V��;-gO.�����Zb�w��
�(	�����+~/iq����f�X����w����p@�'?�#��o���_I�n�w�j���b��6+�b��x�h�o
<�8���8��n��Q�i4ω^[�mNl�v��T�ѣ[ع�6�kw�>�/�������g{+�m-�s�O�&��2���'�d���� ��p���*ŏ�����eᥳ������G��Mbj�R�ڛH��u��A�������_��O�k��W�P���F�jN5�����Np��]5%�wV����NOٓ���x�T���c�����G��op�C�R9s�Er��Ko������ƺ_�?�Ac�iQ\����km��f
[0Ą��	"����F**��_��;~k����G�_��D�/ʋwlc�I���iiD?5�=[��Du��3�D��H
y��*O�W+���E�^���-m�i ���� T�=�};�������u i��QEi7��oɽ�
�8��=�pW�K�Y��k�|�Ĺ�\;/т�}���|[��Ww��B��m}w>TҘ�*���g#���bq>ے^D:|���^6L|>����������~�x��I��s�R��Ik�:*�x�ㅙ[������F�ʲWM�”��w�zg�5���I�i����⸴说xu��-�ϒf���� g�?�f�2��l���f��9\7=
d���� V���ip+�����:�k���y��d��W����%un�-�i�j�,�6)4vlO%n4�w�n O�W�ڿ����{M�����1_ԧ����W��`K�����q��_�@3���Ҿ��������^H�v�p?�l���a<g<ds�=k�
OE���R�M����1���]��U�V�w�3.x9r�_�U��Ǯ:S
�<f�~
�=�h��{r
L��Gu{��=��z�i�׆U�/���d�Zi_�K��*��k��#�o���+���%Z���t��	9��s2�6h\�d�Wy`��q��}|�7/�j�����o�x��[��
�n��_v��z��
��	��5l��� �;�;Kfw�y�Z�(1���!�&(�*�A��ձ���R���iF�	�m���}l����M.~�[��>���%�G�{�B�*�C���Q]?��m����aP�XAǧ�(���S��J�?�pP�ʂ�2]O��?.����4�-����sf�!m�
!�����w���$a� g#����?o�jP۟�<+�����RG[�Q_��T��1��}?�i��q�����#�l	��:)ǡ8?δ���.Vx�W�䷳>������E_�E"�ܗ�1��]W�?oxf{_��?��)A�
�C��/��k�����o��쵉㘃�ep���)������^>��V*OMO��:ь壹���Oۣş<sk�=_�>�Kխo-�Z�;��6��)7d��z�m|/ഉ#_ij��E�����^��x���%�ß����$��x.F8����>es���I6����_<
sa���tԹ;Zv��u��@b篧Z��|
�jl��_O�^�q�_����"B�����
��g5�:}���Mʭ��E��c>��(�l�ӧ�6Z���mR�qtL{#�$��Lh	J`SXT�<3�I=���>�2�S���N6���ן_C�4���
�S�e���&B�5��O5�z;W㵔Ǭ±K�D��Y6��7pzW�H�һ�p�9y���yO��)���Y~��Eu��-NS��7�'����/�!��8�]��5�^�k���U���>��J�^�'<y��z��Q_R��Pa��Q��r?�m9�d��me�ݲ���_���)�j�=���ף�������oi��U�¯
@ǡk�T��~݃�Vq���?�/��:��'W�<VZ�����9�<�~����W
⟄2���E�ݦ2m� H?�=����N7�Q�W�e��;�y�*�S����ϧ��=��]���}�F��<
�Z����U�+K�EQY�4�͓t[,���(�$��_��8�$�I�^�4�~U���� ^8��U�w�_t�nQ�ϋ5P��i��Ez�����Ht�E|�k�W��j�약6On�v��V/��P���S[���v�z[���>;��֯5|0��������Kw�?�+�������s�s��O�`��
�0>�:��&��:���x���n��o5v�*����#~�f�]h2�O�e?/�X��λp<EF�T�k�]���#�1�,VO6ۼ?�?��뷡�O���6�&���+oiE�B�]:[�i�ϸ�c�z+���]i�%[^@��h�#� Q_e��ǖ���w�]ϱ�੿�c_�릛��j����s��8a�(��z���eky&X�
�
o�gx��4C&[q�BA�2�������8�3�+8?3���>8�w�˿�$�������=h����S��='D��/�,���"�#���N	_��֩�Z������_� :l�[��ɲT,�{��$QEy\������}�))e�O��]���ۏ�j��W�aE����~y_����\���?k�!�eţh6L��s' ����pеi��Y�%�����0���Q_�Q�c�b���P�*�V�{��3F�ϵ^���&P�u<b�+�'�G�T�^�����
C�?<	u���<E������ұ�w��=Gj��'���{����hdm��6�Gӟ�W���ў5ey%��Xz�0�*u�I�Zm�ף���a�i�EY2J��[Q��,`�<����+��$޽�~���R��;����	�ml>%x����m*0I�8QE�&B�����8�P�ZQ�����
endstream
endobj
209 0 obj
<< /Type /Annot
/Subtype /Link
/A 210 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 39.1432 147.7500 151.6432 ]
>>
endobj
210 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-3-new.jpg)
>>
endobj
211 0 obj
<<
/Type /XObject
/Subtype /Image
/Width 150
/Height 150
/ColorSpace /DeviceRGB
/Filter /DCTDecode
/BitsPerComponent 8
/Length 8652>>
stream
����JFIF



��;CREATOR: gd-jpeg v1.0 (using IJG JPEG v62), quality = 90
��C




��C
		

����
"

��







	
���

}
!1AQa"q2���#B��R��$3br�	
%&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz���������������������������������������������������������������������������










	
���

w
!1AQaq"2�B����	#3R�br�
$4�%�&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz��������������������������������������������������������������������������
?�?�*�t=���L��&�s��n���u(>������'w� 9R����A����1����
��'�w�`;���V�/�-ճ��";H�L�����Q�205�<m)�A��Ϩ�tr�{�&��>�Ѩ�-lViE���b�Q]�̟c���m��_��7����� i��)�__���m߲_��7���fW*�=�V��Է+������k��"d��6a���})�c�b@����j�$*�ec-6����.�QE�}g�ӧ��;>�N�j��+�G�3^;soĪ'_-�YU�FC@�<׬���Q���ǍS�D�x��ru="�c�x1���1��~��)ƅzu䮓��~���*��!k�����D���&�Ŕ�-�ڱ�oj�[+�˘�e��t8<�R�.<��Q����1Pꖉ���P�����&�$�����<]8���H?q�-���GvgE�V���8*(��<S����$c�����2QG�'�H��/�#��d��>P�����]����=|u��ys�g�r+�_�+���%�������QJa�]z�Q8�E��
[�V�N�X�1���-̐�%y��8���Fi��cUA�9�.�G�צ��f�N���/&�׶r*��������'������n����Q��Р��+��o�M��K��F��B��8�Aee�`T �����CK 㱩�ϔ"N7/}kۜ����'�U������3�]��QEx���^=��o|-���T��+_<x�����9�WQ���r����*�ǿ��<j��%k�&{i�6�$lXv �ʬX8>��6������	B�!��w�i!�V!J�oJ���;8.�;�v� �����Z�$�D�[a�L�C�\�S�a�d�(&��i�^����Icp�-��8}I}U�չEl���*/�"�����4ho�n6����~��a���Za�:��{�>=)E�Kx���5���I��1������(����$c�����2Q]%@��?�O����9<g��u���+B4��-��o�cny=�Ϳ��
?翋�i���h��m�����#ؗ++����i�������*����f�ע�t���u�^1QSv>������<�,϶������
?翋�i���h�J�#�Rs|�z�.�J��s�(����
xg[��M�V��n�mo�$11R|�c;��9W������?�3_K�\�����{�R�/T]6�����jX�Z2�Cy��K�c
*��n_2����N��)|M��]B(n�].�c6��d�t�x���1�[���'�/���"�"�Š��)������4��o3��N<��^�.���J�uI��mUӮ��iҴmio��w!,y��e{d��ʨ��]�������(��̤]��*
q"�.�9ꝎA�a��ѭR䌴���뭅e�����������?�3G�9��'���o�
#��5�6��?�W���}�	�d��)�"ܱ�\y��C�����y7p��w��O�g��P�j���e
��Fdy�y�j����-������w���M�z�����x�xsG��խ�/�O�J�_s�x+������s�	�=�[��H���}}��x�|�����x�y����[���%6pr2q���Ś�K�~�m�Cs���v��8���aS���'�	]�%����F3�����(���V�;m�����T��	X�����������&t(���1�F?u؀
ȏ���7&�cs���#��k�:*Tc�[�7�''�<��q����-�VѼug��-���,��6�E����]��G�W���Q|W�a�Ś'�����+C�����J���fxnd$��3Z�%��(�8V����
}�Kݿ��-���C��ߊ��,����A	ΉbAG��Ĝ�\�^���XI�TzԡG�2��������o�)��/0�1�����T�?�ߴ�JXx���������O��.P$���'�!(Y#cʓ��eV�]�{�08J���'�������z��.>>��Z�����~���?���-?��W�Z��e�&"� �~��偡8#ڻ#5$|�|-Ji����M~��|��|C�x�+-W�~��X���I����^"�7
�|��#��f�|}o���O����i�c��d�9��>�45��3�e ��}V�p��7��y�/n�q�-O��
��K���x�F�a��L�*/�.yO�0�t-?��W�1^)���=�kuvޤ��_K<�^��Ivٯ�_���������
��L)�����O������_���?�E����vtL�`vj���Q�
�,fUS'��2]:���f���������#ß�(C�;�K�Ϩ
6�
j�L�<�+`��NB玵�v��R�tvO۰$�/�X����������%o�$X|�~"̻}?�n9��-e�;�H��w���Z򚢣6Ӿ�S�.W������h�-�f��
&N�tk#�o�]?�5����A�i	�OG/U]��}��p���_O��#�B�x(�f9��Ե(��Α��_�c�O�?�傄��3S�N���gr����k��;����J�jM29&��6�pXh�o#5���U/�M�?�@��C���-N��Z�B��pӎ?O���Q�4
�Ѹ*2K�s����2�T���t���^�T��S�?_���_��ď�[E��q�^Mr]"��;2�P[yk"�\~�vv/\�������ɧx��>
�ޚUף
�4W*w:�%%tx��S����"\K
��=fq�3�np+���5ь^���cs����W�5����'��#����7���?EԗOѵ��Ae�u�h��h��UV
��� ,
;3��Y�ݏ��S�s'�Zi{��h>"�mZ�
���y?xqL�
8#����g3k6YP~�#��Dr���+)����k)�O�z��(����ټ�d� n���Y
��e1��}G�jê�5���C�޽��O;�?S�I��a)�Fzx�XlCR�m��s�>���J�{���PX'� |%q�.B3��ҾCO]���\0��[�k��b!?���)��┸������ �ސ�����u�����GH[{���%�>6�R��A,K��(��nY��O
�l��k��~k]�5%��Mn�����2��Ս[y�oX����$�'q���t��=E[�ئ�|V]��~#��h֩Bj8�v~Z~
���0i=��Ǜ�j��Q��gӥ_�3H��+/L��d�N	��K��.�Y��V��2#._t�q'x���h�h���͸��N>�qҾc��W[�d�]�	�N���+�+{�?����?�&8?��p?�x%��lt�	�WM�ƺ�ڢ�#�m�Wg��6�R��A$��|ߟ�&ugR���Y�>�]8�u&��c�x�d�y5���R�c��]U�������1�#�vC~�{כjz5��Dj�:m#5�:_i�qȗ��(���{S�I�^	?-?[?�Ҟ"q��.�mݴ�m�!h������W;���2���*.?:�����&o2�+�u�_S�Tח�܆h�6�cq�>�v�qX9ci�DӺ��G�GUH�q1\�O����~���"tȴ� ������<}ۏ���	�&��=e�Q�Ο�)�_䟑E|��(ɩ���S�K�t�_��N�O��&�ĚO��
xVʹ�[q����-er
��yb6�Nx9�l���Ho��H=ό\����)o�6��Sx�OF�j�����F�71�m��۰
���;�73�ŝ�v,��;��\�NVh�ҩ
Jp���>��	�|2|j�H�xɿ��$��O]Z�J��O���H�����Ns���/�t.���֗�<S�q��<�MJ��akc*Rq���Vi����>�_�'W����~3�h��'�O���ù�N$��'��A���_)M�I	܄H?�j1i����è5��7�8rnݻ|����H��~%����Z-��߇�$����wZ��Z����DD��#>j
����=k�q���ʜ���l=�`��f���%`��e4=��?�C_I��W�f[Rym,E,G-略ծ������j��%�����?�jF�_�O��/��j���8�F�+M���m�Ť�Os���N9U0O�V����ϥ}<��JT��s���W^�;��>�_�''�m�1|e�;����Ï�R7��IJ������x���D�ʏ�y#�o���Ȩ�[�#�=��Y.o�N�z���m��;��z�BZ�~�Y~�~'?�׆|�|C�2j6>2�V�Y����D`�|��G̙u%v��+�5?�c�-̯*|^�^X|�����Z��i���5|/6��<���0L�x��sa��I[��а��_�b�Z�Z���N�]u9q�s�l���M7���#�o�e=u�[�hֽ��
���|@�f�� P��|�!����b����}*�0�3��Zڊ��/�xLr
��#'�5ȫbs���.���}O?
�8ڳq�IYw~���.���<|]�]<�G�υ����2���O�A5$��'��V����7����k���Vѹ{���3|�~���M�K��&��,��!;C'����p+���}���m[�WS��M��
�7���	���<k��
��jrY�rxv��,[P��ь����+�	����Z/�V�a^k,��Nٹ��INU��=�-IT��k�s�o�+ã~��Q���ı?��ĬD�>��O��H�����%���"�L98�s{H�5s�����wZz�ܜf��{�(\'�8�����H����<#w���K�jIWi7K���Qհy�@<Q)�Y%qP�՗4��d��9H�iT��uH5U-�B*��*Hpz���*�!-l`�x�O����߶�·���)�CxNr3��5��@N�q���}��b����k��
���S�����C�KM���ҿ^ʲ�m|��hAJ:�V��/S��V.��Ք���d�>�&�,p>OLU��#�(zz�շ�F�7�n�@�R�
��;'�q�?�H��?��Q��\�dgy���	��NWѧ���-�_��ڵ�5��p.���ɬ���5]���2��FEWad}��\�7ΰN�+���ErS���
����L�
�īRx�t�~�=y���g���Px8���E{w���G���!�K�p�����x�@m6�,
ь0Њ��6���֝HE;�}t�}��(bg%+F�<�Ǟ+�-[o��Qv�����x�_���l�X�q�~5�����<��0=YA��5��[�S�����t١�a�-^�h1���1�b�Āx��p�f8Ji+�=�r�
H.Z�ǝ�u��(�+S�s� n��uK�J�B	Q����
fk�d+�IDNӕ�ӑ�לj�V�W�tq���'���X��J򝟟�]G%S�7_;��s��[�����	�-���%Ol�آ��#4�����V�����c�e'��J+)Ns|�>'��#��7w����
�2�W�FÑ�Y����	�A"���_�����N�����	��]O�zզ�g�j�x���\:��¨7���9#���?���V	y��@���]&?�yr���L��mN��*Sn�+��g�Rګ��]52O3�&�R�1f�
Td�=����ǟ�L�?�Cx����d�
b�u��T>�������
������8����_�|�}������N�z��-���Ps��_��]'�S�w����A<S��t��]�H�-z�*�rQk��:���	�5�ʪ��D���
|�,h�_�?��į��W�o���ğ
��gF�����v6� xO�tY�G���~c�_8���V��'��{��]'�W�g�+��a1xy�EKT�Zɿ�O�S�k����Pg���{Pͻډ\@\HІ;@b:d@>�־�����*c�S��l��H><~�a�
�S��St��^�_8~v^�mu�c���U�6�lK��Ib���#������)�
�S��l��I���Q���_��['�W��x��q�Δ�ۖ?�mc�N~��
���A�e�o�+����s�*���+]NŊ����1]G�>7|��9��i�
����	m!���<W^L��4�d�
�o��W����\H�|Ep����'�q��6�>���6�s;i�W��>+
��.zmY.�~���)HgH~X@f$u��^ww��ɧeGp�,������F}�w_��o��Ii�
c�-.�_Rp=�Wy�s~��ڇ�%�'�O<�|J@����q���.�OA`���/�8��w�oX��1M0]�p��-�<��d3c��M{\߶���p��^%#���ۿ�������w�?�Fq��?-��VͲڏ݄����<&����_�G�
h_n�6�On:p����B��n|b�{�{J���
�M��#���/�P�_�,�
�6�kg�#Ҋ�����/���J�
7��7�?�z��l=J+I�-�4+
*)�Ā�@-�98<���}O�<%����6�	�5�w�����򁱢X�����n�!�u��B��zW�V�ۜ���>�
�T�.*
�)%�%�~��"�����s�n�h\vn�J�]�����uM->]WQ��m�m��/4�1¢
�bH
��<.4�Z�Ob��y-�!��ъ����ZB��z�����u_�jsG��K�������
��4����=�2���d*�#���jFN��k`�Q���ӹ�O��?��6�ɼd�q���G����	���l;u��m����S��P�ɉaC���c�
~��9F;��V..���Gk���O
�H�Mu2�t�r�1���Ѷ>��I)��$�i�<~u���6���ZN�}g-������<�2�B��,��n�Vʐ+�X\|��9�uk��v���=�ۭ�v�k�6�WGK�֝ƒ��	��~�"�Mn�9W��2��ܵ�^�t�_z�;aR3���KԛJ��[�2u��?�+��n8>��:\����ĖW������W��?���
ʧ�$��f����ɭ�A#[��Km�A�{%ȱ��
�I$�zz�2�U�Ke8Kx��f���
�x�f�n�_��\�_J�ON��fr�+x��$���Z�
x��
��Q��9��^ �uoq�Yj�۝(��)�iX}��FT;ǞB屻�!�[�xz�&��;R����}V���ݡ�?�R�̈́m_Wn��v��6:�JNz����8ʸ
o60���c���x��2i-���h†;z�����帬�_����ɩ/�7o��%';h��/�!�����L��.h���h�9�t0�~צ�4W����[S?���`
�bY;�z��K/!�Dlw�+�I��,<Z��L����[�DC�4,U�� �9z�5�I
����I|�O�QI�f��ӏ��T���~�)}�cn�3�i���X��WU����϶l)|�٫�Q���RS��z�b�%�ne�8�+��ù��z*k=_�#����E}B�%Q�25�qqt���nex`E�+'$(��Q\m��N���m-uZ=�xfڱ^��(l�����>��|sE�%Y��8W|ɽo�:j%���&?�%τcS�����<˭�|�����@�
�A4Q_W��t�_U���5oCz���^H����E�=�^t*v�7��ޫ��bM��$��^c���X�s���Ҋ+=�ц1N1���M�#ʫJ�9Ij�c�h̖����o�o������G~�u�p�"��v'��O�?�W��
z��8�|���[�>�3�S�jϭ���#"F��vE
Z�M݁���QEy9�a�iRV�n��a]�����
endstream
endobj
212 0 obj
<< /Type /Annot
/Subtype /Link
/A 213 0 R
/Border [0 0 0]
/H /I
/Rect [ 290.1338 634.3336 300.9758 644.2542 ]
>>
endobj
213 0 obj
<< /Type /Action
>>
endobj
214 0 obj
<< /Type /Annot
/Subtype /Link
/A 215 0 R
/Border [0 0 0]
/H /I
/Rect [ 413.1300 610.5241 432.1035 620.4447 ]
>>
endobj
215 0 obj
<< /Type /Action
>>
endobj
216 0 obj
<< /Type /Annot
/Subtype /Link
/A 217 0 R
/Border [0 0 0]
/H /I
/Rect [ 536.1555 586.7146 552.4185 596.6352 ]
>>
endobj
217 0 obj
<< /Type /Action
>>
endobj
218 0 obj
<< /Type /Annot
/Subtype /Link
/A 219 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 419.7712 147.7500 532.2712 ]
>>
endobj
219 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-2-new.jpg)
>>
endobj
220 0 obj
<< /Type /Annot
/Subtype /Link
/A 221 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 39.1432 147.7500 151.6432 ]
>>
endobj
221 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-3-new.jpg)
>>
endobj
222 0 obj
<< /Type /Annot
/Subtype /Link
/A 223 0 R
/Border [0 0 0]
/H /I
/Rect [ 290.1338 634.3336 300.9758 644.2542 ]
>>
endobj
223 0 obj
<< /Type /Action
>>
endobj
224 0 obj
<< /Type /Annot
/Subtype /Link
/A 225 0 R
/Border [0 0 0]
/H /I
/Rect [ 413.1300 610.5241 432.1035 620.4447 ]
>>
endobj
225 0 obj
<< /Type /Action
>>
endobj
226 0 obj
<< /Type /Annot
/Subtype /Link
/A 227 0 R
/Border [0 0 0]
/H /I
/Rect [ 536.1555 586.7146 552.4185 596.6352 ]
>>
endobj
227 0 obj
<< /Type /Action
>>
endobj
228 0 obj
<< /Type /Annot
/Subtype /Link
/A 229 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 419.7712 147.7500 532.2712 ]
>>
endobj
229 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-2-new.jpg)
>>
endobj
230 0 obj
<< /Type /Annot
/Subtype /Link
/A 231 0 R
/Border [0 0 0]
/H /I
/Rect [ 35.2500 39.1432 147.7500 151.6432 ]
>>
endobj
231 0 obj
<< /Type /Action
/S /URI
/URI (http://currents.plos.org/md/files/2012/01/figure-3-new.jpg)
>>
endobj
232 0 obj
<< /Type /Page
/Parent 3 0 R
/Annots [ 234 0 R 236 0 R 238 0 R 240 0 R 242 0 R 244 0 R 246 0 R 248 0 R 250 0 R 252 0 R 254 0 R 256 0 R 258 0 R 260 0 R 262 0 R 264 0 R 266 0 R 268 0 R 270 0 R 272 0 R 274 0 R ]
/Contents 233 0 R
>>
endobj
233 0 obj
<<
/Length 21634 >>
stream

q
15.000 36.362 577.500 740.638 re W n
0.965 0.965 0.965 rg
26.250 699.136 555.000 77.864 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l  f
q
35.250 710.386 537.000 66.614 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 3: Identification of human satellite cells on dystrophic myofibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(Fibre dissection was attempted from a DMD extensor digitorum brevis muscle. Whilst Pax7 immunostaining clearly allows )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(satellite cell identification on healthy human single fibres below the basal lamina \(A\), identification is difficult and ambiguous )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(in DMD fibres \(B\) even at high magnification \(B�, C\).)] TJ ET
Q
BT 26.250 664.915 Td /F4 12.0 Tf  [(Discussion)] TJ ET
BT 26.250 644.961 Td /F1 9.8 Tf  [(Studies on human single myofibres can be performed when muscles can be harvested from tendon to tendon, which is possible )] TJ ET
BT 26.250 633.056 Td /F1 9.8 Tf  [(for very small muscles like abdominal, intercostals and gluteux maximus)] TJ ET
0.267 0.267 0.267 rg
BT 337.255 633.056 Td /F1 9.8 Tf  [( [30] [31])] TJ ET
0.271 0.267 0.267 rg
BT 375.202 633.056 Td /F1 9.8 Tf  [(. However, so far, no satellite cell marker )] TJ ET
BT 26.250 621.151 Td /F1 9.8 Tf  [(characterization has performed on such freshly-isolated human fibres. As muscle biopsies for diagnosis are usually taken from )] TJ ET
BT 26.250 609.246 Td /F1 9.8 Tf  [(muscles such as quadriceps and biceps and as these are usually very small pieces of muscle, we wished to obtain preparations )] TJ ET
BT 26.250 597.342 Td /F1 9.8 Tf  [(of muscle biopsies in which we could unequivocally identify satellite cells under the basal lamina of muscle fibres. The technique )] TJ ET
BT 26.250 585.437 Td /F1 9.8 Tf  [(we have described is a suitable method to characterize human satellite cells in their quiescent state underneath the basal )] TJ ET
BT 26.250 573.532 Td /F1 9.8 Tf  [(lamina.)] TJ ET
BT 26.250 554.127 Td /F1 9.8 Tf  [(To date, most of our knowledge about human satellite cells derives from immunostaining muscle cryosections )] TJ ET
0.267 0.267 0.267 rg
BT 500.929 554.127 Td /F1 9.8 Tf  [([8] [32])] TJ ET
0.271 0.267 0.267 rg
BT 530.744 554.127 Td /F1 9.8 Tf  [(. However, )] TJ ET
BT 26.250 542.223 Td /F1 9.8 Tf  [(it is challenging to distinguish on a transverse muscle section a satellite cell from an interstitial cell, or a myonucleus, particularly )] TJ ET
BT 26.250 530.318 Td /F1 9.8 Tf  [(as both of these have been reported to occasionally express Pax7)] TJ ET
0.267 0.267 0.267 rg
BT 311.320 530.318 Td /F1 9.8 Tf  [( [33])] TJ ET
0.271 0.267 0.267 rg
BT 330.294 530.318 Td /F1 9.8 Tf  [(. This has caused confusion in satellite cell quantification )] TJ ET
BT 26.250 518.413 Td /F1 9.8 Tf  [(and characterization, giving rise to contrasting reports )] TJ ET
0.267 0.267 0.267 rg
BT 260.347 518.413 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 271.190 518.413 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 499.008 Td /F1 9.8 Tf  [(Mouse studies have shown that satellite cells express a mesenchymal marker, the integrin CD 29 )] TJ ET
0.267 0.267 0.267 rg
BT 448.366 499.008 Td /F1 9.8 Tf  [([34])] TJ ET
0.271 0.267 0.267 rg
BT 464.629 499.008 Td /F1 9.8 Tf  [(. Our study shows that )] TJ ET
BT 26.250 487.104 Td /F1 9.8 Tf  [(human satellite cells also express CD29. However, the antibody we used against CD29 identifies human, but not mouse, )] TJ ET
BT 26.250 475.199 Td /F1 9.8 Tf  [(satellite cells. This particular antibody might be a useful marker for detecting donor satellite cells in human/mouse xenografts.)] TJ ET
BT 26.250 455.794 Td /F1 9.8 Tf  [(We cannot exclude the possibility that satellite cells in different human muscles are characterized by different markers and )] TJ ET
BT 26.250 443.889 Td /F1 9.8 Tf  [(future investigations are needed in order to establish whether satellite cells in human muscles are heterogeneous, as has been )] TJ ET
BT 26.250 431.985 Td /F1 9.8 Tf  [(shown in mouse)] TJ ET
0.267 0.267 0.267 rg
BT 96.694 431.985 Td /F1 9.8 Tf  [( [6] [35])] TJ ET
0.271 0.267 0.267 rg
BT 129.220 431.985 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 412.580 Td /F1 9.8 Tf  [(Although the technique that we used was not applicable to DMD muscles, we suspect that, were a DMD biopsy available in the )] TJ ET
BT 26.250 400.675 Td /F1 9.8 Tf  [(early stage of the pathology, when fibres are not so fragile, we would be able to obtain fibre fragments for study. Moreover, for )] TJ ET
BT 26.250 388.770 Td /F1 9.8 Tf  [(disorders such as polymyositis, DMD and myotonic dystrophy, where satellite cell number is known to be altered )] TJ ET
0.267 0.267 0.267 rg
BT 512.833 388.770 Td /F1 9.8 Tf  [([36])] TJ ET
0.271 0.267 0.267 rg
BT 529.096 388.770 Td /F1 9.8 Tf  [(, )] TJ ET
BT 26.250 376.866 Td /F1 9.8 Tf  [(quantification of satellite cells would be informative.)] TJ ET
BT 26.250 357.461 Td /F1 9.8 Tf  [(In summary, we have shown that a clear qualitative and quantitative identification of satellite cells can be performed by analysis )] TJ ET
BT 26.250 345.556 Td /F1 9.8 Tf  [(of human muscle fibre fragments. Further investigations are needed in order to explore the cell biology of human satellite cells.)] TJ ET
BT 26.250 326.151 Td /F4 9.8 Tf  [(Corresponding author:)] TJ ET
BT 26.250 306.747 Td /F1 9.8 Tf  [(Luisa Boldrin, UCL, Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.)] TJ ET
BT 26.250 270.144 Td /F4 12.0 Tf  [(Competing interests)] TJ ET
BT 26.250 250.190 Td /F1 9.8 Tf  [(The authors have declared that no competing interests exist.)] TJ ET
BT 26.250 213.587 Td /F4 12.0 Tf  [(Acknowledgements)] TJ ET
BT 26.250 193.633 Td /F1 9.8 Tf  [(We would like to thank the Biobank of the MRC Centre for Neuromuscular Diseases that provides human muscle cells and )] TJ ET
BT 26.250 181.728 Td /F1 9.8 Tf  [(tissue for research. We would also like to thank Dr Geraldine Edge, Department of Anaesthetics, Stanmore Hospital, London )] TJ ET
BT 26.250 169.824 Td /F1 9.8 Tf  [(and Dr Jan Lehovsky, Department of Orthopaedics, Stanmore Hospital, London for providing us with human muscle samples. )] TJ ET
BT 26.250 157.919 Td /F1 9.8 Tf  [(The Pax7 antibody developed by A. Kawakami was obtained from the Developmental Studies Hybridoma Bank developed )] TJ ET
BT 26.250 146.014 Td /F1 9.8 Tf  [(under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA )] TJ ET
BT 26.250 134.109 Td /F1 9.8 Tf  [(52242.)] TJ ET
BT 26.250 105.007 Td /F4 12.0 Tf  [(References)] TJ ET
BT 26.250 77.553 Td /F1 9.8 Tf  [(1.)] TJ ET
BT 38.132 77.553 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE \(2007\) Activating muscle stem cells: therapeutic potential in muscle diseases. Curr Opin Neurol 20: 577-)] TJ ET
BT 26.250 65.648 Td /F1 9.8 Tf  [(582.)] TJ ET
BT 26.250 46.243 Td /F1 9.8 Tf  [(2.)] TJ ET
BT 38.132 46.243 Td /F1 9.8 Tf  [(Mauro A \(1961\) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9: 493-495.)] TJ ET
Q
q
15.000 36.362 577.500 740.638 re W n
0.965 0.965 0.965 rg
26.250 699.136 555.000 77.864 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l  f
q
35.250 710.386 537.000 66.614 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 3: Identification of human satellite cells on dystrophic myofibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(Fibre dissection was attempted from a DMD extensor digitorum brevis muscle. Whilst Pax7 immunostaining clearly allows )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(satellite cell identification on healthy human single fibres below the basal lamina \(A\), identification is difficult and ambiguous )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(in DMD fibres \(B\) even at high magnification \(B�, C\).)] TJ ET
Q
BT 26.250 664.915 Td /F4 12.0 Tf  [(Discussion)] TJ ET
BT 26.250 644.961 Td /F1 9.8 Tf  [(Studies on human single myofibres can be performed when muscles can be harvested from tendon to tendon, which is possible )] TJ ET
BT 26.250 633.056 Td /F1 9.8 Tf  [(for very small muscles like abdominal, intercostals and gluteux maximus)] TJ ET
0.267 0.267 0.267 rg
BT 337.255 633.056 Td /F1 9.8 Tf  [( [30] [31])] TJ ET
0.271 0.267 0.267 rg
BT 375.202 633.056 Td /F1 9.8 Tf  [(. However, so far, no satellite cell marker )] TJ ET
BT 26.250 621.151 Td /F1 9.8 Tf  [(characterization has performed on such freshly-isolated human fibres. As muscle biopsies for diagnosis are usually taken from )] TJ ET
BT 26.250 609.246 Td /F1 9.8 Tf  [(muscles such as quadriceps and biceps and as these are usually very small pieces of muscle, we wished to obtain preparations )] TJ ET
BT 26.250 597.342 Td /F1 9.8 Tf  [(of muscle biopsies in which we could unequivocally identify satellite cells under the basal lamina of muscle fibres. The technique )] TJ ET
BT 26.250 585.437 Td /F1 9.8 Tf  [(we have described is a suitable method to characterize human satellite cells in their quiescent state underneath the basal )] TJ ET
BT 26.250 573.532 Td /F1 9.8 Tf  [(lamina.)] TJ ET
BT 26.250 554.127 Td /F1 9.8 Tf  [(To date, most of our knowledge about human satellite cells derives from immunostaining muscle cryosections )] TJ ET
0.267 0.267 0.267 rg
BT 500.929 554.127 Td /F1 9.8 Tf  [([8] [32])] TJ ET
0.271 0.267 0.267 rg
BT 530.744 554.127 Td /F1 9.8 Tf  [(. However, )] TJ ET
BT 26.250 542.223 Td /F1 9.8 Tf  [(it is challenging to distinguish on a transverse muscle section a satellite cell from an interstitial cell, or a myonucleus, particularly )] TJ ET
BT 26.250 530.318 Td /F1 9.8 Tf  [(as both of these have been reported to occasionally express Pax7)] TJ ET
0.267 0.267 0.267 rg
BT 311.320 530.318 Td /F1 9.8 Tf  [( [33])] TJ ET
0.271 0.267 0.267 rg
BT 330.294 530.318 Td /F1 9.8 Tf  [(. This has caused confusion in satellite cell quantification )] TJ ET
BT 26.250 518.413 Td /F1 9.8 Tf  [(and characterization, giving rise to contrasting reports )] TJ ET
0.267 0.267 0.267 rg
BT 260.347 518.413 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 271.190 518.413 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 499.008 Td /F1 9.8 Tf  [(Mouse studies have shown that satellite cells express a mesenchymal marker, the integrin CD 29 )] TJ ET
0.267 0.267 0.267 rg
BT 448.366 499.008 Td /F1 9.8 Tf  [([34])] TJ ET
0.271 0.267 0.267 rg
BT 464.629 499.008 Td /F1 9.8 Tf  [(. Our study shows that )] TJ ET
BT 26.250 487.104 Td /F1 9.8 Tf  [(human satellite cells also express CD29. However, the antibody we used against CD29 identifies human, but not mouse, )] TJ ET
BT 26.250 475.199 Td /F1 9.8 Tf  [(satellite cells. This particular antibody might be a useful marker for detecting donor satellite cells in human/mouse xenografts.)] TJ ET
BT 26.250 455.794 Td /F1 9.8 Tf  [(We cannot exclude the possibility that satellite cells in different human muscles are characterized by different markers and )] TJ ET
BT 26.250 443.889 Td /F1 9.8 Tf  [(future investigations are needed in order to establish whether satellite cells in human muscles are heterogeneous, as has been )] TJ ET
BT 26.250 431.985 Td /F1 9.8 Tf  [(shown in mouse)] TJ ET
0.267 0.267 0.267 rg
BT 96.694 431.985 Td /F1 9.8 Tf  [( [6] [35])] TJ ET
0.271 0.267 0.267 rg
BT 129.220 431.985 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 412.580 Td /F1 9.8 Tf  [(Although the technique that we used was not applicable to DMD muscles, we suspect that, were a DMD biopsy available in the )] TJ ET
BT 26.250 400.675 Td /F1 9.8 Tf  [(early stage of the pathology, when fibres are not so fragile, we would be able to obtain fibre fragments for study. Moreover, for )] TJ ET
BT 26.250 388.770 Td /F1 9.8 Tf  [(disorders such as polymyositis, DMD and myotonic dystrophy, where satellite cell number is known to be altered )] TJ ET
0.267 0.267 0.267 rg
BT 512.833 388.770 Td /F1 9.8 Tf  [([36])] TJ ET
0.271 0.267 0.267 rg
BT 529.096 388.770 Td /F1 9.8 Tf  [(, )] TJ ET
BT 26.250 376.866 Td /F1 9.8 Tf  [(quantification of satellite cells would be informative.)] TJ ET
BT 26.250 357.461 Td /F1 9.8 Tf  [(In summary, we have shown that a clear qualitative and quantitative identification of satellite cells can be performed by analysis )] TJ ET
BT 26.250 345.556 Td /F1 9.8 Tf  [(of human muscle fibre fragments. Further investigations are needed in order to explore the cell biology of human satellite cells.)] TJ ET
BT 26.250 326.151 Td /F4 9.8 Tf  [(Corresponding author:)] TJ ET
BT 26.250 306.747 Td /F1 9.8 Tf  [(Luisa Boldrin, UCL, Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.)] TJ ET
BT 26.250 270.144 Td /F4 12.0 Tf  [(Competing interests)] TJ ET
BT 26.250 250.190 Td /F1 9.8 Tf  [(The authors have declared that no competing interests exist.)] TJ ET
BT 26.250 213.587 Td /F4 12.0 Tf  [(Acknowledgements)] TJ ET
BT 26.250 193.633 Td /F1 9.8 Tf  [(We would like to thank the Biobank of the MRC Centre for Neuromuscular Diseases that provides human muscle cells and )] TJ ET
BT 26.250 181.728 Td /F1 9.8 Tf  [(tissue for research. We would also like to thank Dr Geraldine Edge, Department of Anaesthetics, Stanmore Hospital, London )] TJ ET
BT 26.250 169.824 Td /F1 9.8 Tf  [(and Dr Jan Lehovsky, Department of Orthopaedics, Stanmore Hospital, London for providing us with human muscle samples. )] TJ ET
BT 26.250 157.919 Td /F1 9.8 Tf  [(The Pax7 antibody developed by A. Kawakami was obtained from the Developmental Studies Hybridoma Bank developed )] TJ ET
BT 26.250 146.014 Td /F1 9.8 Tf  [(under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA )] TJ ET
BT 26.250 134.109 Td /F1 9.8 Tf  [(52242.)] TJ ET
BT 26.250 105.007 Td /F4 12.0 Tf  [(References)] TJ ET
BT 26.250 77.553 Td /F1 9.8 Tf  [(1.)] TJ ET
BT 38.132 77.553 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE \(2007\) Activating muscle stem cells: therapeutic potential in muscle diseases. Curr Opin Neurol 20: 577-)] TJ ET
BT 26.250 65.648 Td /F1 9.8 Tf  [(582.)] TJ ET
BT 26.250 46.243 Td /F1 9.8 Tf  [(2.)] TJ ET
BT 38.132 46.243 Td /F1 9.8 Tf  [(Mauro A \(1961\) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9: 493-495.)] TJ ET
Q
q
15.000 36.362 577.500 740.638 re W n
0.965 0.965 0.965 rg
26.250 699.136 555.000 77.864 re f
0.267 0.267 0.267 rg
0.267 0.267 0.267 RG
26.250 699.136 m 581.250 699.136 l 581.250 699.886 l 26.250 699.886 l  f
q
35.250 710.386 537.000 66.614 re W n
0.271 0.267 0.267 rg
BT 35.250 767.476 Td /F4 9.8 Tf  [(Fig. 3: Identification of human satellite cells on dystrophic myofibres.)] TJ ET
BT 35.250 748.106 Td /F1 9.8 Tf  [(Fibre dissection was attempted from a DMD extensor digitorum brevis muscle. Whilst Pax7 immunostaining clearly allows )] TJ ET
BT 35.250 734.370 Td /F1 9.8 Tf  [(satellite cell identification on healthy human single fibres below the basal lamina \(A\), identification is difficult and ambiguous )] TJ ET
BT 35.250 720.634 Td /F1 9.8 Tf  [(in DMD fibres \(B\) even at high magnification \(B�, C\).)] TJ ET
Q
BT 26.250 664.915 Td /F4 12.0 Tf  [(Discussion)] TJ ET
BT 26.250 644.961 Td /F1 9.8 Tf  [(Studies on human single myofibres can be performed when muscles can be harvested from tendon to tendon, which is possible )] TJ ET
BT 26.250 633.056 Td /F1 9.8 Tf  [(for very small muscles like abdominal, intercostals and gluteux maximus)] TJ ET
0.267 0.267 0.267 rg
BT 337.255 633.056 Td /F1 9.8 Tf  [( [30] [31])] TJ ET
0.271 0.267 0.267 rg
BT 375.202 633.056 Td /F1 9.8 Tf  [(. However, so far, no satellite cell marker )] TJ ET
BT 26.250 621.151 Td /F1 9.8 Tf  [(characterization has performed on such freshly-isolated human fibres. As muscle biopsies for diagnosis are usually taken from )] TJ ET
BT 26.250 609.246 Td /F1 9.8 Tf  [(muscles such as quadriceps and biceps and as these are usually very small pieces of muscle, we wished to obtain preparations )] TJ ET
BT 26.250 597.342 Td /F1 9.8 Tf  [(of muscle biopsies in which we could unequivocally identify satellite cells under the basal lamina of muscle fibres. The technique )] TJ ET
BT 26.250 585.437 Td /F1 9.8 Tf  [(we have described is a suitable method to characterize human satellite cells in their quiescent state underneath the basal )] TJ ET
BT 26.250 573.532 Td /F1 9.8 Tf  [(lamina.)] TJ ET
BT 26.250 554.127 Td /F1 9.8 Tf  [(To date, most of our knowledge about human satellite cells derives from immunostaining muscle cryosections )] TJ ET
0.267 0.267 0.267 rg
BT 500.929 554.127 Td /F1 9.8 Tf  [([8] [32])] TJ ET
0.271 0.267 0.267 rg
BT 530.744 554.127 Td /F1 9.8 Tf  [(. However, )] TJ ET
BT 26.250 542.223 Td /F1 9.8 Tf  [(it is challenging to distinguish on a transverse muscle section a satellite cell from an interstitial cell, or a myonucleus, particularly )] TJ ET
BT 26.250 530.318 Td /F1 9.8 Tf  [(as both of these have been reported to occasionally express Pax7)] TJ ET
0.267 0.267 0.267 rg
BT 311.320 530.318 Td /F1 9.8 Tf  [( [33])] TJ ET
0.271 0.267 0.267 rg
BT 330.294 530.318 Td /F1 9.8 Tf  [(. This has caused confusion in satellite cell quantification )] TJ ET
BT 26.250 518.413 Td /F1 9.8 Tf  [(and characterization, giving rise to contrasting reports )] TJ ET
0.267 0.267 0.267 rg
BT 260.347 518.413 Td /F1 9.8 Tf  [([8])] TJ ET
0.271 0.267 0.267 rg
BT 271.190 518.413 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 499.008 Td /F1 9.8 Tf  [(Mouse studies have shown that satellite cells express a mesenchymal marker, the integrin CD 29 )] TJ ET
0.267 0.267 0.267 rg
BT 448.366 499.008 Td /F1 9.8 Tf  [([34])] TJ ET
0.271 0.267 0.267 rg
BT 464.629 499.008 Td /F1 9.8 Tf  [(. Our study shows that )] TJ ET
BT 26.250 487.104 Td /F1 9.8 Tf  [(human satellite cells also express CD29. However, the antibody we used against CD29 identifies human, but not mouse, )] TJ ET
BT 26.250 475.199 Td /F1 9.8 Tf  [(satellite cells. This particular antibody might be a useful marker for detecting donor satellite cells in human/mouse xenografts.)] TJ ET
BT 26.250 455.794 Td /F1 9.8 Tf  [(We cannot exclude the possibility that satellite cells in different human muscles are characterized by different markers and )] TJ ET
BT 26.250 443.889 Td /F1 9.8 Tf  [(future investigations are needed in order to establish whether satellite cells in human muscles are heterogeneous, as has been )] TJ ET
BT 26.250 431.985 Td /F1 9.8 Tf  [(shown in mouse)] TJ ET
0.267 0.267 0.267 rg
BT 96.694 431.985 Td /F1 9.8 Tf  [( [6] [35])] TJ ET
0.271 0.267 0.267 rg
BT 129.220 431.985 Td /F1 9.8 Tf  [(.)] TJ ET
BT 26.250 412.580 Td /F1 9.8 Tf  [(Although the technique that we used was not applicable to DMD muscles, we suspect that, were a DMD biopsy available in the )] TJ ET
BT 26.250 400.675 Td /F1 9.8 Tf  [(early stage of the pathology, when fibres are not so fragile, we would be able to obtain fibre fragments for study. Moreover, for )] TJ ET
BT 26.250 388.770 Td /F1 9.8 Tf  [(disorders such as polymyositis, DMD and myotonic dystrophy, where satellite cell number is known to be altered )] TJ ET
0.267 0.267 0.267 rg
BT 512.833 388.770 Td /F1 9.8 Tf  [([36])] TJ ET
0.271 0.267 0.267 rg
BT 529.096 388.770 Td /F1 9.8 Tf  [(, )] TJ ET
BT 26.250 376.866 Td /F1 9.8 Tf  [(quantification of satellite cells would be informative.)] TJ ET
BT 26.250 357.461 Td /F1 9.8 Tf  [(In summary, we have shown that a clear qualitative and quantitative identification of satellite cells can be performed by analysis )] TJ ET
BT 26.250 345.556 Td /F1 9.8 Tf  [(of human muscle fibre fragments. Further investigations are needed in order to explore the cell biology of human satellite cells.)] TJ ET
BT 26.250 326.151 Td /F4 9.8 Tf  [(Corresponding author:)] TJ ET
BT 26.250 306.747 Td /F1 9.8 Tf  [(Luisa Boldrin, UCL, Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.)] TJ ET
BT 26.250 270.144 Td /F4 12.0 Tf  [(Competing interests)] TJ ET
BT 26.250 250.190 Td /F1 9.8 Tf  [(The authors have declared that no competing interests exist.)] TJ ET
BT 26.250 213.587 Td /F4 12.0 Tf  [(Acknowledgements)] TJ ET
BT 26.250 193.633 Td /F1 9.8 Tf  [(We would like to thank the Biobank of the MRC Centre for Neuromuscular Diseases that provides human muscle cells and )] TJ ET
BT 26.250 181.728 Td /F1 9.8 Tf  [(tissue for research. We would also like to thank Dr Geraldine Edge, Department of Anaesthetics, Stanmore Hospital, London )] TJ ET
BT 26.250 169.824 Td /F1 9.8 Tf  [(and Dr Jan Lehovsky, Department of Orthopaedics, Stanmore Hospital, London for providing us with human muscle samples. )] TJ ET
BT 26.250 157.919 Td /F1 9.8 Tf  [(The Pax7 antibody developed by A. Kawakami was obtained from the Developmental Studies Hybridoma Bank developed )] TJ ET
BT 26.250 146.014 Td /F1 9.8 Tf  [(under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA )] TJ ET
BT 26.250 134.109 Td /F1 9.8 Tf  [(52242.)] TJ ET
BT 26.250 105.007 Td /F4 12.0 Tf  [(References)] TJ ET
BT 26.250 77.553 Td /F1 9.8 Tf  [(1.)] TJ ET
BT 38.132 77.553 Td /F1 9.8 Tf  [(Boldrin L, Morgan JE \(2007\) Activating muscle stem cells: therapeutic potential in muscle diseases. Curr Opin Neurol 20: 577-)] TJ ET
BT 26.250 65.648 Td /F1 9.8 Tf  [(582.)] TJ ET
BT 26.250 46.243 Td /F1 9.8 Tf  [(2.)] TJ ET
BT 38.132 46.243 Td /F1 9.8 Tf  [(Mauro A \(1961\) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9: 493-495.)] TJ ET
Q
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(4)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
234 0 obj
<< /Type /Annot
/Subtype /Link
/A 235 0 R
/Border [0 0 0]
/H /I
/Rect [ 337.2555 632.1541 375.2025 642.0747 ]
>>
endobj
235 0 obj
<< /Type /Action
>>
endobj
236 0 obj
<< /Type /Annot
/Subtype /Link
/A 237 0 R
/Border [0 0 0]
/H /I
/Rect [ 500.9287 553.2256 530.7442 563.1462 ]
>>
endobj
237 0 obj
<< /Type /Action
>>
endobj
238 0 obj
<< /Type /Annot
/Subtype /Link
/A 239 0 R
/Border [0 0 0]
/H /I
/Rect [ 311.3205 529.4161 330.2940 539.3367 ]
>>
endobj
239 0 obj
<< /Type /Action
>>
endobj
240 0 obj
<< /Type /Annot
/Subtype /Link
/A 241 0 R
/Border [0 0 0]
/H /I
/Rect [ 260.3475 517.5113 271.1895 527.4319 ]
>>
endobj
241 0 obj
<< /Type /Action
>>
endobj
242 0 obj
<< /Type /Annot
/Subtype /Link
/A 243 0 R
/Border [0 0 0]
/H /I
/Rect [ 448.3665 498.1066 464.6295 508.0272 ]
>>
endobj
243 0 obj
<< /Type /Action
>>
endobj
244 0 obj
<< /Type /Annot
/Subtype /Link
/A 245 0 R
/Border [0 0 0]
/H /I
/Rect [ 96.6937 431.0828 129.2198 441.0034 ]
>>
endobj
245 0 obj
<< /Type /Action
>>
endobj
246 0 obj
<< /Type /Annot
/Subtype /Link
/A 247 0 R
/Border [0 0 0]
/H /I
/Rect [ 512.8335 387.8686 529.0965 397.7892 ]
>>
endobj
247 0 obj
<< /Type /Action
>>
endobj
248 0 obj
<< /Type /Annot
/Subtype /Link
/A 249 0 R
/Border [0 0 0]
/H /I
/Rect [ 337.2555 632.1541 375.2025 642.0747 ]
>>
endobj
249 0 obj
<< /Type /Action
>>
endobj
250 0 obj
<< /Type /Annot
/Subtype /Link
/A 251 0 R
/Border [0 0 0]
/H /I
/Rect [ 500.9287 553.2256 530.7442 563.1462 ]
>>
endobj
251 0 obj
<< /Type /Action
>>
endobj
252 0 obj
<< /Type /Annot
/Subtype /Link
/A 253 0 R
/Border [0 0 0]
/H /I
/Rect [ 311.3205 529.4161 330.2940 539.3367 ]
>>
endobj
253 0 obj
<< /Type /Action
>>
endobj
254 0 obj
<< /Type /Annot
/Subtype /Link
/A 255 0 R
/Border [0 0 0]
/H /I
/Rect [ 260.3475 517.5113 271.1895 527.4319 ]
>>
endobj
255 0 obj
<< /Type /Action
>>
endobj
256 0 obj
<< /Type /Annot
/Subtype /Link
/A 257 0 R
/Border [0 0 0]
/H /I
/Rect [ 448.3665 498.1066 464.6295 508.0272 ]
>>
endobj
257 0 obj
<< /Type /Action
>>
endobj
258 0 obj
<< /Type /Annot
/Subtype /Link
/A 259 0 R
/Border [0 0 0]
/H /I
/Rect [ 96.6937 431.0828 129.2198 441.0034 ]
>>
endobj
259 0 obj
<< /Type /Action
>>
endobj
260 0 obj
<< /Type /Annot
/Subtype /Link
/A 261 0 R
/Border [0 0 0]
/H /I
/Rect [ 512.8335 387.8686 529.0965 397.7892 ]
>>
endobj
261 0 obj
<< /Type /Action
>>
endobj
262 0 obj
<< /Type /Annot
/Subtype /Link
/A 263 0 R
/Border [0 0 0]
/H /I
/Rect [ 337.2555 632.1541 375.2025 642.0747 ]
>>
endobj
263 0 obj
<< /Type /Action
>>
endobj
264 0 obj
<< /Type /Annot
/Subtype /Link
/A 265 0 R
/Border [0 0 0]
/H /I
/Rect [ 500.9287 553.2256 530.7442 563.1462 ]
>>
endobj
265 0 obj
<< /Type /Action
>>
endobj
266 0 obj
<< /Type /Annot
/Subtype /Link
/A 267 0 R
/Border [0 0 0]
/H /I
/Rect [ 311.3205 529.4161 330.2940 539.3367 ]
>>
endobj
267 0 obj
<< /Type /Action
>>
endobj
268 0 obj
<< /Type /Annot
/Subtype /Link
/A 269 0 R
/Border [0 0 0]
/H /I
/Rect [ 260.3475 517.5113 271.1895 527.4319 ]
>>
endobj
269 0 obj
<< /Type /Action
>>
endobj
270 0 obj
<< /Type /Annot
/Subtype /Link
/A 271 0 R
/Border [0 0 0]
/H /I
/Rect [ 448.3665 498.1066 464.6295 508.0272 ]
>>
endobj
271 0 obj
<< /Type /Action
>>
endobj
272 0 obj
<< /Type /Annot
/Subtype /Link
/A 273 0 R
/Border [0 0 0]
/H /I
/Rect [ 96.6937 431.0828 129.2198 441.0034 ]
>>
endobj
273 0 obj
<< /Type /Action
>>
endobj
274 0 obj
<< /Type /Annot
/Subtype /Link
/A 275 0 R
/Border [0 0 0]
/H /I
/Rect [ 512.8335 387.8686 529.0965 397.7892 ]
>>
endobj
275 0 obj
<< /Type /Action
>>
endobj
276 0 obj
<< /Type /Page
/Parent 3 0 R
/Contents 277 0 R
>>
endobj
277 0 obj
<<
/Length 22048 >>
stream

0.271 0.267 0.267 rg
q
15.000 41.881 577.500 735.119 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(3.)] TJ ET
BT 38.132 759.976 Td /F1 9.8 Tf  [(Rosenblatt JD, Lunt AI, Parry DJ, Partridge TA \(1995\) Culturing satellite cells from living single muscle fiber explants. In Vitro )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Cell Dev Biol Anim 31: 773-779.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(4.)] TJ ET
BT 38.132 728.667 Td /F1 9.8 Tf  [(Bischoff R \(1975\) Regeneration of single skeletal muscle fibers in vitro. Anat Rec 182: 215-235.)] TJ ET
BT 26.250 709.262 Td /F1 9.8 Tf  [(5.)] TJ ET
BT 38.132 709.262 Td /F1 9.8 Tf  [(Boldrin L, Zammit PS, Muntoni F, Morgan JE \(2009\) Mature adult dystrophic mouse muscle environment does not impede )] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(efficient engrafted satellite cell regeneration and self-renewal. Stem Cells 27: 2478-2487.)] TJ ET
BT 26.250 677.952 Td /F1 9.8 Tf  [(6.)] TJ ET
BT 38.132 677.952 Td /F1 9.8 Tf  [(Collins CA, Partridge TA \(2005\) Self-Renewal of the Adult Skeletal Muscle Satellite Cell. Cell Cycle 4.)] TJ ET
BT 26.250 658.548 Td /F1 9.8 Tf  [(7.)] TJ ET
BT 38.132 658.548 Td /F1 9.8 Tf  [(Collins CA, Zammit PS, Ruiz AP, Morgan JE, Partridge TA \(2007\) A population of myogenic stem cells that survives skeletal )] TJ ET
BT 26.250 646.643 Td /F1 9.8 Tf  [(muscle aging. Stem Cells 25: 885-894.)] TJ ET
BT 26.250 627.238 Td /F1 9.8 Tf  [(8.)] TJ ET
BT 38.132 627.238 Td /F1 9.8 Tf  [(Boldrin L, Muntoni F, Morgan JE \(2010\) Are human and mouse satellite cells really the same? The journal of histochemistry )] TJ ET
BT 26.250 615.333 Td /F1 9.8 Tf  [(and cytochemistry : official journal of the Histochemistry Society 58: 941-955.)] TJ ET
BT 26.250 595.929 Td /F1 9.8 Tf  [(9.)] TJ ET
BT 38.132 595.929 Td /F1 9.8 Tf  [(Seale P, Sabourin LA, Girgis-Gabardo A, Mansouri A, Gruss P, et al. \(2000\) Pax7 is required for the specification of )] TJ ET
BT 26.250 584.024 Td /F1 9.8 Tf  [(myogenic satellite cells. Cell 102: 777-786.)] TJ ET
BT 26.250 564.619 Td /F1 9.8 Tf  [(10.)] TJ ET
BT 43.553 564.619 Td /F1 9.8 Tf  [(Irintchev A, Zeschnigk M, Starzinski-Powitz A, Wernig A \(1994\) Expression pattern of M-cadherin in normal, denervated, )] TJ ET
BT 26.250 552.714 Td /F1 9.8 Tf  [(and regenerating mouse muscles. Dev Dyn 199: 326-337.)] TJ ET
BT 26.250 533.310 Td /F1 9.8 Tf  [(11.)] TJ ET
BT 43.553 533.310 Td /F1 9.8 Tf  [(Beauchamp JR, Heslop L, Yu DS, Tajbakhsh S, Kelly RG, et al. \(2000\) Expression of CD34 and Myf5 defines the majority )] TJ ET
BT 26.250 521.405 Td /F1 9.8 Tf  [(of quiescent adult skeletal muscle satellite cells. J Cell Biol 151: 1221-1234.)] TJ ET
BT 26.250 502.000 Td /F1 9.8 Tf  [(12.)] TJ ET
BT 43.553 502.000 Td /F1 9.8 Tf  [(Volonte D, Liu Y, Galbiati F \(2005\) The modulation of caveolin-1 expression controls satellite cell activation during muscle )] TJ ET
BT 26.250 490.095 Td /F1 9.8 Tf  [(repair. FASEB J 19: 237-239.)] TJ ET
BT 26.250 470.691 Td /F1 9.8 Tf  [(13.)] TJ ET
BT 43.553 470.691 Td /F1 9.8 Tf  [(Gnocchi VF, White RB, Ono Y, Ellis JA, Zammit PS \(2009\) Further characterisation of the molecular signature of quiescent )] TJ ET
BT 26.250 458.786 Td /F1 9.8 Tf  [(and activated mouse muscle satellite cells. PLoS One 4: e5205.)] TJ ET
BT 26.250 439.381 Td /F1 9.8 Tf  [(14.)] TJ ET
BT 43.553 439.381 Td /F1 9.8 Tf  [(Fukada S, Uezumi A, Ikemoto M, Masuda S, Segawa M, et al. \(2007\) Molecular signature of quiescent satellite cells in adult )] TJ ET
BT 26.250 427.476 Td /F1 9.8 Tf  [(skeletal muscle. Stem Cells 25: 2448-2459.)] TJ ET
BT 26.250 408.072 Td /F1 9.8 Tf  [(15.)] TJ ET
BT 43.553 408.072 Td /F1 9.8 Tf  [(Day K, Shefer G, Richardson JB, Enikolopov G, Yablonka-Reuveni Z \(2007\) Nestin-GFP reporter expression defines the )] TJ ET
BT 26.250 396.167 Td /F1 9.8 Tf  [(quiescent state of skeletal muscle satellite cells. Dev Biol 304: 246-259.)] TJ ET
BT 26.250 376.762 Td /F1 9.8 Tf  [(16.)] TJ ET
BT 43.553 376.762 Td /F1 9.8 Tf  [(Blanco-Bose WE, Yao CC, Kramer RH, Blau HM \(2001\) Purification of mouse primary myoblasts based on alpha 7 integrin )] TJ ET
BT 26.250 364.857 Td /F1 9.8 Tf  [(expression. Exp Cell Res 265: 212-220.)] TJ ET
BT 26.250 345.453 Td /F1 9.8 Tf  [(17.)] TJ ET
BT 43.553 345.453 Td /F1 9.8 Tf  [(Zammit PS, Golding JP, Nagata Y, Hudon V, Partridge TA, et al. \(2004\) Muscle satellite cells adopt divergent fates: a )] TJ ET
BT 26.250 333.548 Td /F1 9.8 Tf  [(mechanism for self-renewal? J Cell Biol 166: 347-357.)] TJ ET
BT 26.250 314.143 Td /F1 9.8 Tf  [(18.)] TJ ET
BT 43.553 314.143 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Day K, Vine A, Shefer G \(2008\) Defining the transcriptional signature of skeletal muscle stem cells. J )] TJ ET
BT 26.250 302.238 Td /F1 9.8 Tf  [(Anim Sci 86: E207-216.)] TJ ET
BT 26.250 282.834 Td /F1 9.8 Tf  [(19.)] TJ ET
BT 43.553 282.834 Td /F1 9.8 Tf  [(Grounds MD, Garrett KL, Lai MC, Wright WE, Beilharz MW \(1992\) Identification of skeletal muscle precursor cells in vivo by )] TJ ET
BT 26.250 270.929 Td /F1 9.8 Tf  [(use of MyoD1 and myogenin probes. Cell Tissue Res 267: 99-104.)] TJ ET
BT 26.250 251.524 Td /F1 9.8 Tf  [(20.)] TJ ET
BT 43.553 251.524 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Rivera AJ \(1994\) Temporal expression of regulatory and structural muscle proteins during myogenesis )] TJ ET
BT 26.250 239.619 Td /F1 9.8 Tf  [(of satellite cells on isolated adult rat fibers. Dev Biol 164: 588-603.)] TJ ET
BT 26.250 220.215 Td /F1 9.8 Tf  [(21.)] TJ ET
BT 43.553 220.215 Td /F1 9.8 Tf  [(Collins CA, Olsen I, Zammit PS, Heslop L, Petrie A, et al. \(2005\) Stem cell function, self-renewal, and behavioral )] TJ ET
BT 26.250 208.310 Td /F1 9.8 Tf  [(heterogeneity of cells from the adult muscle satellite cell niche. Cell 122: 289-301.)] TJ ET
BT 26.250 188.905 Td /F1 9.8 Tf  [(22.)] TJ ET
BT 43.553 188.905 Td /F1 9.8 Tf  [(Kuang S, Kuroda K, Le Grand F, Rudnicki MA \(2007\) Asymmetric self-renewal and commitment of satellite stem cells in )] TJ ET
BT 26.250 177.000 Td /F1 9.8 Tf  [(muscle. Cell 129: 999-1010.)] TJ ET
BT 26.250 157.596 Td /F1 9.8 Tf  [(23.)] TJ ET
BT 43.553 157.596 Td /F1 9.8 Tf  [(Sacco A, Doyonnas R, Kraft P, Vitorovic S, Blau HM \(2008\) Self-renewal and expansion of single transplanted muscle stem )] TJ ET
BT 26.250 145.691 Td /F1 9.8 Tf  [(cells. Nature 456: 502-506.)] TJ ET
BT 26.250 126.286 Td /F1 9.8 Tf  [(24.)] TJ ET
BT 43.553 126.286 Td /F1 9.8 Tf  [(Boldrin L, Zammit P, Muntoni F, Morgan J \(2009\) The Mature Adult Dystrophic Mouse Muscle Environment Does Not )] TJ ET
BT 26.250 114.381 Td /F1 9.8 Tf  [(Impede Efficient Engrafted Satellite Cell Regeneration And Self-Renewal. Stem Cells.)] TJ ET
BT 26.250 94.977 Td /F1 9.8 Tf  [(25.)] TJ ET
BT 43.553 94.977 Td /F1 9.8 Tf  [(Kelly R, Alonso S, Tajbakhsh S, Cossu G, Buckingham M \(1995\) Myosin light chain 3F regulatory sequences confer )] TJ ET
BT 26.250 83.072 Td /F1 9.8 Tf  [(regionalized cardiac and skeletal muscle expression in transgenic mice. J Cell Biol 129: 383-396.)] TJ ET
BT 26.250 63.667 Td /F1 9.8 Tf  [(26.)] TJ ET
BT 43.553 63.667 Td /F1 9.8 Tf  [(Valdez MR, Richardson JA, Klein WH, Olson EN \(2000\) Failure of Myf5 to support myogenic differentiation without )] TJ ET
BT 26.250 51.762 Td /F1 9.8 Tf  [(myogenin, MyoD, and MRF4. Dev Biol 219: 287-298.)] TJ ET
Q
q
15.000 41.881 577.500 735.119 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(3.)] TJ ET
BT 38.132 759.976 Td /F1 9.8 Tf  [(Rosenblatt JD, Lunt AI, Parry DJ, Partridge TA \(1995\) Culturing satellite cells from living single muscle fiber explants. In Vitro )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Cell Dev Biol Anim 31: 773-779.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(4.)] TJ ET
BT 38.132 728.667 Td /F1 9.8 Tf  [(Bischoff R \(1975\) Regeneration of single skeletal muscle fibers in vitro. Anat Rec 182: 215-235.)] TJ ET
BT 26.250 709.262 Td /F1 9.8 Tf  [(5.)] TJ ET
BT 38.132 709.262 Td /F1 9.8 Tf  [(Boldrin L, Zammit PS, Muntoni F, Morgan JE \(2009\) Mature adult dystrophic mouse muscle environment does not impede )] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(efficient engrafted satellite cell regeneration and self-renewal. Stem Cells 27: 2478-2487.)] TJ ET
BT 26.250 677.952 Td /F1 9.8 Tf  [(6.)] TJ ET
BT 38.132 677.952 Td /F1 9.8 Tf  [(Collins CA, Partridge TA \(2005\) Self-Renewal of the Adult Skeletal Muscle Satellite Cell. Cell Cycle 4.)] TJ ET
BT 26.250 658.548 Td /F1 9.8 Tf  [(7.)] TJ ET
BT 38.132 658.548 Td /F1 9.8 Tf  [(Collins CA, Zammit PS, Ruiz AP, Morgan JE, Partridge TA \(2007\) A population of myogenic stem cells that survives skeletal )] TJ ET
BT 26.250 646.643 Td /F1 9.8 Tf  [(muscle aging. Stem Cells 25: 885-894.)] TJ ET
BT 26.250 627.238 Td /F1 9.8 Tf  [(8.)] TJ ET
BT 38.132 627.238 Td /F1 9.8 Tf  [(Boldrin L, Muntoni F, Morgan JE \(2010\) Are human and mouse satellite cells really the same? The journal of histochemistry )] TJ ET
BT 26.250 615.333 Td /F1 9.8 Tf  [(and cytochemistry : official journal of the Histochemistry Society 58: 941-955.)] TJ ET
BT 26.250 595.929 Td /F1 9.8 Tf  [(9.)] TJ ET
BT 38.132 595.929 Td /F1 9.8 Tf  [(Seale P, Sabourin LA, Girgis-Gabardo A, Mansouri A, Gruss P, et al. \(2000\) Pax7 is required for the specification of )] TJ ET
BT 26.250 584.024 Td /F1 9.8 Tf  [(myogenic satellite cells. Cell 102: 777-786.)] TJ ET
BT 26.250 564.619 Td /F1 9.8 Tf  [(10.)] TJ ET
BT 43.553 564.619 Td /F1 9.8 Tf  [(Irintchev A, Zeschnigk M, Starzinski-Powitz A, Wernig A \(1994\) Expression pattern of M-cadherin in normal, denervated, )] TJ ET
BT 26.250 552.714 Td /F1 9.8 Tf  [(and regenerating mouse muscles. Dev Dyn 199: 326-337.)] TJ ET
BT 26.250 533.310 Td /F1 9.8 Tf  [(11.)] TJ ET
BT 43.553 533.310 Td /F1 9.8 Tf  [(Beauchamp JR, Heslop L, Yu DS, Tajbakhsh S, Kelly RG, et al. \(2000\) Expression of CD34 and Myf5 defines the majority )] TJ ET
BT 26.250 521.405 Td /F1 9.8 Tf  [(of quiescent adult skeletal muscle satellite cells. J Cell Biol 151: 1221-1234.)] TJ ET
BT 26.250 502.000 Td /F1 9.8 Tf  [(12.)] TJ ET
BT 43.553 502.000 Td /F1 9.8 Tf  [(Volonte D, Liu Y, Galbiati F \(2005\) The modulation of caveolin-1 expression controls satellite cell activation during muscle )] TJ ET
BT 26.250 490.095 Td /F1 9.8 Tf  [(repair. FASEB J 19: 237-239.)] TJ ET
BT 26.250 470.691 Td /F1 9.8 Tf  [(13.)] TJ ET
BT 43.553 470.691 Td /F1 9.8 Tf  [(Gnocchi VF, White RB, Ono Y, Ellis JA, Zammit PS \(2009\) Further characterisation of the molecular signature of quiescent )] TJ ET
BT 26.250 458.786 Td /F1 9.8 Tf  [(and activated mouse muscle satellite cells. PLoS One 4: e5205.)] TJ ET
BT 26.250 439.381 Td /F1 9.8 Tf  [(14.)] TJ ET
BT 43.553 439.381 Td /F1 9.8 Tf  [(Fukada S, Uezumi A, Ikemoto M, Masuda S, Segawa M, et al. \(2007\) Molecular signature of quiescent satellite cells in adult )] TJ ET
BT 26.250 427.476 Td /F1 9.8 Tf  [(skeletal muscle. Stem Cells 25: 2448-2459.)] TJ ET
BT 26.250 408.072 Td /F1 9.8 Tf  [(15.)] TJ ET
BT 43.553 408.072 Td /F1 9.8 Tf  [(Day K, Shefer G, Richardson JB, Enikolopov G, Yablonka-Reuveni Z \(2007\) Nestin-GFP reporter expression defines the )] TJ ET
BT 26.250 396.167 Td /F1 9.8 Tf  [(quiescent state of skeletal muscle satellite cells. Dev Biol 304: 246-259.)] TJ ET
BT 26.250 376.762 Td /F1 9.8 Tf  [(16.)] TJ ET
BT 43.553 376.762 Td /F1 9.8 Tf  [(Blanco-Bose WE, Yao CC, Kramer RH, Blau HM \(2001\) Purification of mouse primary myoblasts based on alpha 7 integrin )] TJ ET
BT 26.250 364.857 Td /F1 9.8 Tf  [(expression. Exp Cell Res 265: 212-220.)] TJ ET
BT 26.250 345.453 Td /F1 9.8 Tf  [(17.)] TJ ET
BT 43.553 345.453 Td /F1 9.8 Tf  [(Zammit PS, Golding JP, Nagata Y, Hudon V, Partridge TA, et al. \(2004\) Muscle satellite cells adopt divergent fates: a )] TJ ET
BT 26.250 333.548 Td /F1 9.8 Tf  [(mechanism for self-renewal? J Cell Biol 166: 347-357.)] TJ ET
BT 26.250 314.143 Td /F1 9.8 Tf  [(18.)] TJ ET
BT 43.553 314.143 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Day K, Vine A, Shefer G \(2008\) Defining the transcriptional signature of skeletal muscle stem cells. J )] TJ ET
BT 26.250 302.238 Td /F1 9.8 Tf  [(Anim Sci 86: E207-216.)] TJ ET
BT 26.250 282.834 Td /F1 9.8 Tf  [(19.)] TJ ET
BT 43.553 282.834 Td /F1 9.8 Tf  [(Grounds MD, Garrett KL, Lai MC, Wright WE, Beilharz MW \(1992\) Identification of skeletal muscle precursor cells in vivo by )] TJ ET
BT 26.250 270.929 Td /F1 9.8 Tf  [(use of MyoD1 and myogenin probes. Cell Tissue Res 267: 99-104.)] TJ ET
BT 26.250 251.524 Td /F1 9.8 Tf  [(20.)] TJ ET
BT 43.553 251.524 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Rivera AJ \(1994\) Temporal expression of regulatory and structural muscle proteins during myogenesis )] TJ ET
BT 26.250 239.619 Td /F1 9.8 Tf  [(of satellite cells on isolated adult rat fibers. Dev Biol 164: 588-603.)] TJ ET
BT 26.250 220.215 Td /F1 9.8 Tf  [(21.)] TJ ET
BT 43.553 220.215 Td /F1 9.8 Tf  [(Collins CA, Olsen I, Zammit PS, Heslop L, Petrie A, et al. \(2005\) Stem cell function, self-renewal, and behavioral )] TJ ET
BT 26.250 208.310 Td /F1 9.8 Tf  [(heterogeneity of cells from the adult muscle satellite cell niche. Cell 122: 289-301.)] TJ ET
BT 26.250 188.905 Td /F1 9.8 Tf  [(22.)] TJ ET
BT 43.553 188.905 Td /F1 9.8 Tf  [(Kuang S, Kuroda K, Le Grand F, Rudnicki MA \(2007\) Asymmetric self-renewal and commitment of satellite stem cells in )] TJ ET
BT 26.250 177.000 Td /F1 9.8 Tf  [(muscle. Cell 129: 999-1010.)] TJ ET
BT 26.250 157.596 Td /F1 9.8 Tf  [(23.)] TJ ET
BT 43.553 157.596 Td /F1 9.8 Tf  [(Sacco A, Doyonnas R, Kraft P, Vitorovic S, Blau HM \(2008\) Self-renewal and expansion of single transplanted muscle stem )] TJ ET
BT 26.250 145.691 Td /F1 9.8 Tf  [(cells. Nature 456: 502-506.)] TJ ET
BT 26.250 126.286 Td /F1 9.8 Tf  [(24.)] TJ ET
BT 43.553 126.286 Td /F1 9.8 Tf  [(Boldrin L, Zammit P, Muntoni F, Morgan J \(2009\) The Mature Adult Dystrophic Mouse Muscle Environment Does Not )] TJ ET
BT 26.250 114.381 Td /F1 9.8 Tf  [(Impede Efficient Engrafted Satellite Cell Regeneration And Self-Renewal. Stem Cells.)] TJ ET
BT 26.250 94.977 Td /F1 9.8 Tf  [(25.)] TJ ET
BT 43.553 94.977 Td /F1 9.8 Tf  [(Kelly R, Alonso S, Tajbakhsh S, Cossu G, Buckingham M \(1995\) Myosin light chain 3F regulatory sequences confer )] TJ ET
BT 26.250 83.072 Td /F1 9.8 Tf  [(regionalized cardiac and skeletal muscle expression in transgenic mice. J Cell Biol 129: 383-396.)] TJ ET
BT 26.250 63.667 Td /F1 9.8 Tf  [(26.)] TJ ET
BT 43.553 63.667 Td /F1 9.8 Tf  [(Valdez MR, Richardson JA, Klein WH, Olson EN \(2000\) Failure of Myf5 to support myogenic differentiation without )] TJ ET
BT 26.250 51.762 Td /F1 9.8 Tf  [(myogenin, MyoD, and MRF4. Dev Biol 219: 287-298.)] TJ ET
Q
q
15.000 41.881 577.500 735.119 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(3.)] TJ ET
BT 38.132 759.976 Td /F1 9.8 Tf  [(Rosenblatt JD, Lunt AI, Parry DJ, Partridge TA \(1995\) Culturing satellite cells from living single muscle fiber explants. In Vitro )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(Cell Dev Biol Anim 31: 773-779.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(4.)] TJ ET
BT 38.132 728.667 Td /F1 9.8 Tf  [(Bischoff R \(1975\) Regeneration of single skeletal muscle fibers in vitro. Anat Rec 182: 215-235.)] TJ ET
BT 26.250 709.262 Td /F1 9.8 Tf  [(5.)] TJ ET
BT 38.132 709.262 Td /F1 9.8 Tf  [(Boldrin L, Zammit PS, Muntoni F, Morgan JE \(2009\) Mature adult dystrophic mouse muscle environment does not impede )] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(efficient engrafted satellite cell regeneration and self-renewal. Stem Cells 27: 2478-2487.)] TJ ET
BT 26.250 677.952 Td /F1 9.8 Tf  [(6.)] TJ ET
BT 38.132 677.952 Td /F1 9.8 Tf  [(Collins CA, Partridge TA \(2005\) Self-Renewal of the Adult Skeletal Muscle Satellite Cell. Cell Cycle 4.)] TJ ET
BT 26.250 658.548 Td /F1 9.8 Tf  [(7.)] TJ ET
BT 38.132 658.548 Td /F1 9.8 Tf  [(Collins CA, Zammit PS, Ruiz AP, Morgan JE, Partridge TA \(2007\) A population of myogenic stem cells that survives skeletal )] TJ ET
BT 26.250 646.643 Td /F1 9.8 Tf  [(muscle aging. Stem Cells 25: 885-894.)] TJ ET
BT 26.250 627.238 Td /F1 9.8 Tf  [(8.)] TJ ET
BT 38.132 627.238 Td /F1 9.8 Tf  [(Boldrin L, Muntoni F, Morgan JE \(2010\) Are human and mouse satellite cells really the same? The journal of histochemistry )] TJ ET
BT 26.250 615.333 Td /F1 9.8 Tf  [(and cytochemistry : official journal of the Histochemistry Society 58: 941-955.)] TJ ET
BT 26.250 595.929 Td /F1 9.8 Tf  [(9.)] TJ ET
BT 38.132 595.929 Td /F1 9.8 Tf  [(Seale P, Sabourin LA, Girgis-Gabardo A, Mansouri A, Gruss P, et al. \(2000\) Pax7 is required for the specification of )] TJ ET
BT 26.250 584.024 Td /F1 9.8 Tf  [(myogenic satellite cells. Cell 102: 777-786.)] TJ ET
BT 26.250 564.619 Td /F1 9.8 Tf  [(10.)] TJ ET
BT 43.553 564.619 Td /F1 9.8 Tf  [(Irintchev A, Zeschnigk M, Starzinski-Powitz A, Wernig A \(1994\) Expression pattern of M-cadherin in normal, denervated, )] TJ ET
BT 26.250 552.714 Td /F1 9.8 Tf  [(and regenerating mouse muscles. Dev Dyn 199: 326-337.)] TJ ET
BT 26.250 533.310 Td /F1 9.8 Tf  [(11.)] TJ ET
BT 43.553 533.310 Td /F1 9.8 Tf  [(Beauchamp JR, Heslop L, Yu DS, Tajbakhsh S, Kelly RG, et al. \(2000\) Expression of CD34 and Myf5 defines the majority )] TJ ET
BT 26.250 521.405 Td /F1 9.8 Tf  [(of quiescent adult skeletal muscle satellite cells. J Cell Biol 151: 1221-1234.)] TJ ET
BT 26.250 502.000 Td /F1 9.8 Tf  [(12.)] TJ ET
BT 43.553 502.000 Td /F1 9.8 Tf  [(Volonte D, Liu Y, Galbiati F \(2005\) The modulation of caveolin-1 expression controls satellite cell activation during muscle )] TJ ET
BT 26.250 490.095 Td /F1 9.8 Tf  [(repair. FASEB J 19: 237-239.)] TJ ET
BT 26.250 470.691 Td /F1 9.8 Tf  [(13.)] TJ ET
BT 43.553 470.691 Td /F1 9.8 Tf  [(Gnocchi VF, White RB, Ono Y, Ellis JA, Zammit PS \(2009\) Further characterisation of the molecular signature of quiescent )] TJ ET
BT 26.250 458.786 Td /F1 9.8 Tf  [(and activated mouse muscle satellite cells. PLoS One 4: e5205.)] TJ ET
BT 26.250 439.381 Td /F1 9.8 Tf  [(14.)] TJ ET
BT 43.553 439.381 Td /F1 9.8 Tf  [(Fukada S, Uezumi A, Ikemoto M, Masuda S, Segawa M, et al. \(2007\) Molecular signature of quiescent satellite cells in adult )] TJ ET
BT 26.250 427.476 Td /F1 9.8 Tf  [(skeletal muscle. Stem Cells 25: 2448-2459.)] TJ ET
BT 26.250 408.072 Td /F1 9.8 Tf  [(15.)] TJ ET
BT 43.553 408.072 Td /F1 9.8 Tf  [(Day K, Shefer G, Richardson JB, Enikolopov G, Yablonka-Reuveni Z \(2007\) Nestin-GFP reporter expression defines the )] TJ ET
BT 26.250 396.167 Td /F1 9.8 Tf  [(quiescent state of skeletal muscle satellite cells. Dev Biol 304: 246-259.)] TJ ET
BT 26.250 376.762 Td /F1 9.8 Tf  [(16.)] TJ ET
BT 43.553 376.762 Td /F1 9.8 Tf  [(Blanco-Bose WE, Yao CC, Kramer RH, Blau HM \(2001\) Purification of mouse primary myoblasts based on alpha 7 integrin )] TJ ET
BT 26.250 364.857 Td /F1 9.8 Tf  [(expression. Exp Cell Res 265: 212-220.)] TJ ET
BT 26.250 345.453 Td /F1 9.8 Tf  [(17.)] TJ ET
BT 43.553 345.453 Td /F1 9.8 Tf  [(Zammit PS, Golding JP, Nagata Y, Hudon V, Partridge TA, et al. \(2004\) Muscle satellite cells adopt divergent fates: a )] TJ ET
BT 26.250 333.548 Td /F1 9.8 Tf  [(mechanism for self-renewal? J Cell Biol 166: 347-357.)] TJ ET
BT 26.250 314.143 Td /F1 9.8 Tf  [(18.)] TJ ET
BT 43.553 314.143 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Day K, Vine A, Shefer G \(2008\) Defining the transcriptional signature of skeletal muscle stem cells. J )] TJ ET
BT 26.250 302.238 Td /F1 9.8 Tf  [(Anim Sci 86: E207-216.)] TJ ET
BT 26.250 282.834 Td /F1 9.8 Tf  [(19.)] TJ ET
BT 43.553 282.834 Td /F1 9.8 Tf  [(Grounds MD, Garrett KL, Lai MC, Wright WE, Beilharz MW \(1992\) Identification of skeletal muscle precursor cells in vivo by )] TJ ET
BT 26.250 270.929 Td /F1 9.8 Tf  [(use of MyoD1 and myogenin probes. Cell Tissue Res 267: 99-104.)] TJ ET
BT 26.250 251.524 Td /F1 9.8 Tf  [(20.)] TJ ET
BT 43.553 251.524 Td /F1 9.8 Tf  [(Yablonka-Reuveni Z, Rivera AJ \(1994\) Temporal expression of regulatory and structural muscle proteins during myogenesis )] TJ ET
BT 26.250 239.619 Td /F1 9.8 Tf  [(of satellite cells on isolated adult rat fibers. Dev Biol 164: 588-603.)] TJ ET
BT 26.250 220.215 Td /F1 9.8 Tf  [(21.)] TJ ET
BT 43.553 220.215 Td /F1 9.8 Tf  [(Collins CA, Olsen I, Zammit PS, Heslop L, Petrie A, et al. \(2005\) Stem cell function, self-renewal, and behavioral )] TJ ET
BT 26.250 208.310 Td /F1 9.8 Tf  [(heterogeneity of cells from the adult muscle satellite cell niche. Cell 122: 289-301.)] TJ ET
BT 26.250 188.905 Td /F1 9.8 Tf  [(22.)] TJ ET
BT 43.553 188.905 Td /F1 9.8 Tf  [(Kuang S, Kuroda K, Le Grand F, Rudnicki MA \(2007\) Asymmetric self-renewal and commitment of satellite stem cells in )] TJ ET
BT 26.250 177.000 Td /F1 9.8 Tf  [(muscle. Cell 129: 999-1010.)] TJ ET
BT 26.250 157.596 Td /F1 9.8 Tf  [(23.)] TJ ET
BT 43.553 157.596 Td /F1 9.8 Tf  [(Sacco A, Doyonnas R, Kraft P, Vitorovic S, Blau HM \(2008\) Self-renewal and expansion of single transplanted muscle stem )] TJ ET
BT 26.250 145.691 Td /F1 9.8 Tf  [(cells. Nature 456: 502-506.)] TJ ET
BT 26.250 126.286 Td /F1 9.8 Tf  [(24.)] TJ ET
BT 43.553 126.286 Td /F1 9.8 Tf  [(Boldrin L, Zammit P, Muntoni F, Morgan J \(2009\) The Mature Adult Dystrophic Mouse Muscle Environment Does Not )] TJ ET
BT 26.250 114.381 Td /F1 9.8 Tf  [(Impede Efficient Engrafted Satellite Cell Regeneration And Self-Renewal. Stem Cells.)] TJ ET
BT 26.250 94.977 Td /F1 9.8 Tf  [(25.)] TJ ET
BT 43.553 94.977 Td /F1 9.8 Tf  [(Kelly R, Alonso S, Tajbakhsh S, Cossu G, Buckingham M \(1995\) Myosin light chain 3F regulatory sequences confer )] TJ ET
BT 26.250 83.072 Td /F1 9.8 Tf  [(regionalized cardiac and skeletal muscle expression in transgenic mice. J Cell Biol 129: 383-396.)] TJ ET
BT 26.250 63.667 Td /F1 9.8 Tf  [(26.)] TJ ET
BT 43.553 63.667 Td /F1 9.8 Tf  [(Valdez MR, Richardson JA, Klein WH, Olson EN \(2000\) Failure of Myf5 to support myogenic differentiation without )] TJ ET
BT 26.250 51.762 Td /F1 9.8 Tf  [(myogenin, MyoD, and MRF4. Dev Biol 219: 287-298.)] TJ ET
Q
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(5)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
278 0 obj
<< /Type /Page
/Parent 3 0 R
/Contents 279 0 R
>>
endobj
279 0 obj
<<
/Length 9739 >>
stream

0.271 0.267 0.267 rg
q
15.000 448.905 577.500 328.095 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(27.)] TJ ET
BT 43.553 759.976 Td /F1 9.8 Tf  [(Rando TA \(2001\) The dystrophin-glycoprotein complex, cellular signaling, and the regulation of cell survival in the muscular )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(dystrophies. Muscle Nerve 24: 1575-1594.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(28.)] TJ ET
BT 43.553 728.667 Td /F1 9.8 Tf  [(Poelstra KA, Eijkelkamp MF, Veldhuizen AG \(2000\) The geometry of the human paraspinal muscles with the aid of three-)] TJ ET
BT 26.250 716.762 Td /F1 9.8 Tf  [(dimensional computed tomography scans and 3-Space Isotrak. Spine 25: 2176-2179.)] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(29.)] TJ ET
BT 43.553 697.357 Td /F1 9.8 Tf  [(Burkholder TJ \(2001\) Age does not influence muscle fiber length adaptation to increased excursion. Journal of applied )] TJ ET
BT 26.250 685.452 Td /F1 9.8 Tf  [(physiology 91: 2466-2470.)] TJ ET
BT 26.250 666.048 Td /F1 9.8 Tf  [(30.)] TJ ET
BT 43.553 666.048 Td /F1 9.8 Tf  [(Bonavaud S, Agbulut O, D'Honneur G, Nizard R, Mouly V, et al. \(2002\) Preparation of isolated human muscle fibers: a )] TJ ET
BT 26.250 654.143 Td /F1 9.8 Tf  [(technical report. In Vitro Cell Dev Biol Anim 38: 66-72.)] TJ ET
BT 26.250 634.738 Td /F1 9.8 Tf  [(31.)] TJ ET
BT 43.553 634.738 Td /F1 9.8 Tf  [(De Coppi P, Milan G, Scarda A, Boldrin L, Centobene C, et al. \(2006\) Rosiglitazone modifies the adipogenic potential of )] TJ ET
BT 26.250 622.833 Td /F1 9.8 Tf  [(human muscle satellite cells. Diabetologia 49: 1962-1973.)] TJ ET
BT 26.250 603.429 Td /F1 9.8 Tf  [(32.)] TJ ET
BT 43.553 603.429 Td /F1 9.8 Tf  [(Lindstrom M, Thornell LE \(2009\) New multiple labelling method for improved satellite cell identification in human muscle: )] TJ ET
BT 26.250 591.524 Td /F1 9.8 Tf  [(application to a cohort of power-lifters and sedentary men. Histochem Cell Biol 132: 141-157.)] TJ ET
BT 26.250 572.119 Td /F1 9.8 Tf  [(33.)] TJ ET
BT 43.553 572.119 Td /F1 9.8 Tf  [(Reimann J, Brimah K, Schroder R, Wernig A, Beauchamp JR, et al. \(2004\) Pax7 distribution in human skeletal muscle )] TJ ET
BT 26.250 560.214 Td /F1 9.8 Tf  [(biopsies and myogenic tissue cultures. Cell Tissue Res 315: 233-242.)] TJ ET
BT 26.250 540.810 Td /F1 9.8 Tf  [(34.)] TJ ET
BT 43.553 540.810 Td /F1 9.8 Tf  [(Bosnakovski D, Xu Z, Li W, Thet S, Cleaver O, et al. \(2008\) Prospective isolation of skeletal muscle stem cells with a Pax7 )] TJ ET
BT 26.250 528.905 Td /F1 9.8 Tf  [(reporter. Stem Cells 26: 3194-3204.)] TJ ET
BT 26.250 509.500 Td /F1 9.8 Tf  [(35.)] TJ ET
BT 43.553 509.500 Td /F1 9.8 Tf  [(Ono Y, Boldrin L, Knopp P, Morgan JE, Zammit PS \(2009\) Muscle satellite cells are a functionally heterogeneous population )] TJ ET
BT 26.250 497.595 Td /F1 9.8 Tf  [(in both somite-derived and branchiomeric muscles. Dev Biol 337: 29-41.)] TJ ET
BT 26.250 478.191 Td /F1 9.8 Tf  [(36.)] TJ ET
BT 43.553 478.191 Td /F1 9.8 Tf  [(Ishimoto S, Goto I, Ohta M, Kuroiwa Y \(1983\) A quantitative study of the muscle satellite cells in various neuromuscular )] TJ ET
BT 26.250 466.286 Td /F1 9.8 Tf  [(disorders. J Neurol Sci 62: 303-314.)] TJ ET
Q
q
15.000 448.905 577.500 328.095 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(27.)] TJ ET
BT 43.553 759.976 Td /F1 9.8 Tf  [(Rando TA \(2001\) The dystrophin-glycoprotein complex, cellular signaling, and the regulation of cell survival in the muscular )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(dystrophies. Muscle Nerve 24: 1575-1594.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(28.)] TJ ET
BT 43.553 728.667 Td /F1 9.8 Tf  [(Poelstra KA, Eijkelkamp MF, Veldhuizen AG \(2000\) The geometry of the human paraspinal muscles with the aid of three-)] TJ ET
BT 26.250 716.762 Td /F1 9.8 Tf  [(dimensional computed tomography scans and 3-Space Isotrak. Spine 25: 2176-2179.)] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(29.)] TJ ET
BT 43.553 697.357 Td /F1 9.8 Tf  [(Burkholder TJ \(2001\) Age does not influence muscle fiber length adaptation to increased excursion. Journal of applied )] TJ ET
BT 26.250 685.452 Td /F1 9.8 Tf  [(physiology 91: 2466-2470.)] TJ ET
BT 26.250 666.048 Td /F1 9.8 Tf  [(30.)] TJ ET
BT 43.553 666.048 Td /F1 9.8 Tf  [(Bonavaud S, Agbulut O, D'Honneur G, Nizard R, Mouly V, et al. \(2002\) Preparation of isolated human muscle fibers: a )] TJ ET
BT 26.250 654.143 Td /F1 9.8 Tf  [(technical report. In Vitro Cell Dev Biol Anim 38: 66-72.)] TJ ET
BT 26.250 634.738 Td /F1 9.8 Tf  [(31.)] TJ ET
BT 43.553 634.738 Td /F1 9.8 Tf  [(De Coppi P, Milan G, Scarda A, Boldrin L, Centobene C, et al. \(2006\) Rosiglitazone modifies the adipogenic potential of )] TJ ET
BT 26.250 622.833 Td /F1 9.8 Tf  [(human muscle satellite cells. Diabetologia 49: 1962-1973.)] TJ ET
BT 26.250 603.429 Td /F1 9.8 Tf  [(32.)] TJ ET
BT 43.553 603.429 Td /F1 9.8 Tf  [(Lindstrom M, Thornell LE \(2009\) New multiple labelling method for improved satellite cell identification in human muscle: )] TJ ET
BT 26.250 591.524 Td /F1 9.8 Tf  [(application to a cohort of power-lifters and sedentary men. Histochem Cell Biol 132: 141-157.)] TJ ET
BT 26.250 572.119 Td /F1 9.8 Tf  [(33.)] TJ ET
BT 43.553 572.119 Td /F1 9.8 Tf  [(Reimann J, Brimah K, Schroder R, Wernig A, Beauchamp JR, et al. \(2004\) Pax7 distribution in human skeletal muscle )] TJ ET
BT 26.250 560.214 Td /F1 9.8 Tf  [(biopsies and myogenic tissue cultures. Cell Tissue Res 315: 233-242.)] TJ ET
BT 26.250 540.810 Td /F1 9.8 Tf  [(34.)] TJ ET
BT 43.553 540.810 Td /F1 9.8 Tf  [(Bosnakovski D, Xu Z, Li W, Thet S, Cleaver O, et al. \(2008\) Prospective isolation of skeletal muscle stem cells with a Pax7 )] TJ ET
BT 26.250 528.905 Td /F1 9.8 Tf  [(reporter. Stem Cells 26: 3194-3204.)] TJ ET
BT 26.250 509.500 Td /F1 9.8 Tf  [(35.)] TJ ET
BT 43.553 509.500 Td /F1 9.8 Tf  [(Ono Y, Boldrin L, Knopp P, Morgan JE, Zammit PS \(2009\) Muscle satellite cells are a functionally heterogeneous population )] TJ ET
BT 26.250 497.595 Td /F1 9.8 Tf  [(in both somite-derived and branchiomeric muscles. Dev Biol 337: 29-41.)] TJ ET
BT 26.250 478.191 Td /F1 9.8 Tf  [(36.)] TJ ET
BT 43.553 478.191 Td /F1 9.8 Tf  [(Ishimoto S, Goto I, Ohta M, Kuroiwa Y \(1983\) A quantitative study of the muscle satellite cells in various neuromuscular )] TJ ET
BT 26.250 466.286 Td /F1 9.8 Tf  [(disorders. J Neurol Sci 62: 303-314.)] TJ ET
Q
q
15.000 448.905 577.500 328.095 re W n
0.271 0.267 0.267 rg
BT 26.250 759.976 Td /F1 9.8 Tf  [(27.)] TJ ET
BT 43.553 759.976 Td /F1 9.8 Tf  [(Rando TA \(2001\) The dystrophin-glycoprotein complex, cellular signaling, and the regulation of cell survival in the muscular )] TJ ET
BT 26.250 748.071 Td /F1 9.8 Tf  [(dystrophies. Muscle Nerve 24: 1575-1594.)] TJ ET
BT 26.250 728.667 Td /F1 9.8 Tf  [(28.)] TJ ET
BT 43.553 728.667 Td /F1 9.8 Tf  [(Poelstra KA, Eijkelkamp MF, Veldhuizen AG \(2000\) The geometry of the human paraspinal muscles with the aid of three-)] TJ ET
BT 26.250 716.762 Td /F1 9.8 Tf  [(dimensional computed tomography scans and 3-Space Isotrak. Spine 25: 2176-2179.)] TJ ET
BT 26.250 697.357 Td /F1 9.8 Tf  [(29.)] TJ ET
BT 43.553 697.357 Td /F1 9.8 Tf  [(Burkholder TJ \(2001\) Age does not influence muscle fiber length adaptation to increased excursion. Journal of applied )] TJ ET
BT 26.250 685.452 Td /F1 9.8 Tf  [(physiology 91: 2466-2470.)] TJ ET
BT 26.250 666.048 Td /F1 9.8 Tf  [(30.)] TJ ET
BT 43.553 666.048 Td /F1 9.8 Tf  [(Bonavaud S, Agbulut O, D'Honneur G, Nizard R, Mouly V, et al. \(2002\) Preparation of isolated human muscle fibers: a )] TJ ET
BT 26.250 654.143 Td /F1 9.8 Tf  [(technical report. In Vitro Cell Dev Biol Anim 38: 66-72.)] TJ ET
BT 26.250 634.738 Td /F1 9.8 Tf  [(31.)] TJ ET
BT 43.553 634.738 Td /F1 9.8 Tf  [(De Coppi P, Milan G, Scarda A, Boldrin L, Centobene C, et al. \(2006\) Rosiglitazone modifies the adipogenic potential of )] TJ ET
BT 26.250 622.833 Td /F1 9.8 Tf  [(human muscle satellite cells. Diabetologia 49: 1962-1973.)] TJ ET
BT 26.250 603.429 Td /F1 9.8 Tf  [(32.)] TJ ET
BT 43.553 603.429 Td /F1 9.8 Tf  [(Lindstrom M, Thornell LE \(2009\) New multiple labelling method for improved satellite cell identification in human muscle: )] TJ ET
BT 26.250 591.524 Td /F1 9.8 Tf  [(application to a cohort of power-lifters and sedentary men. Histochem Cell Biol 132: 141-157.)] TJ ET
BT 26.250 572.119 Td /F1 9.8 Tf  [(33.)] TJ ET
BT 43.553 572.119 Td /F1 9.8 Tf  [(Reimann J, Brimah K, Schroder R, Wernig A, Beauchamp JR, et al. \(2004\) Pax7 distribution in human skeletal muscle )] TJ ET
BT 26.250 560.214 Td /F1 9.8 Tf  [(biopsies and myogenic tissue cultures. Cell Tissue Res 315: 233-242.)] TJ ET
BT 26.250 540.810 Td /F1 9.8 Tf  [(34.)] TJ ET
BT 43.553 540.810 Td /F1 9.8 Tf  [(Bosnakovski D, Xu Z, Li W, Thet S, Cleaver O, et al. \(2008\) Prospective isolation of skeletal muscle stem cells with a Pax7 )] TJ ET
BT 26.250 528.905 Td /F1 9.8 Tf  [(reporter. Stem Cells 26: 3194-3204.)] TJ ET
BT 26.250 509.500 Td /F1 9.8 Tf  [(35.)] TJ ET
BT 43.553 509.500 Td /F1 9.8 Tf  [(Ono Y, Boldrin L, Knopp P, Morgan JE, Zammit PS \(2009\) Muscle satellite cells are a functionally heterogeneous population )] TJ ET
BT 26.250 497.595 Td /F1 9.8 Tf  [(in both somite-derived and branchiomeric muscles. Dev Biol 337: 29-41.)] TJ ET
BT 26.250 478.191 Td /F1 9.8 Tf  [(36.)] TJ ET
BT 43.553 478.191 Td /F1 9.8 Tf  [(Ishimoto S, Goto I, Ohta M, Kuroiwa Y \(1983\) A quantitative study of the muscle satellite cells in various neuromuscular )] TJ ET
BT 26.250 466.286 Td /F1 9.8 Tf  [(disorders. J Neurol Sci 62: 303-314.)] TJ ET
Q
q
0.000 0.000 0.000 rg
BT 291.710 19.825 Td /F1 11.0 Tf  [(6)] TJ ET
BT 25.000 19.825 Td /F1 11.0 Tf  [(PLOS Currents Muscular Dystrophy)] TJ ET

Q
endstream
endobj
xref
0 280
0000000000 65535 f 
0000000008 00000 n 
0000000073 00000 n 
0000000119 00000 n 
0000000431 00000 n 
0000000468 00000 n 
0000000688 00000 n 
0000001248 00000 n 
0000028438 00000 n 
0000028545 00000 n 
0000028653 00000 n 
0000028764 00000 n 
0000028877 00000 n 
0000029265 00000 n 
0000033622 00000 n 
0000033749 00000 n 
0000033893 00000 n 
0000034019 00000 n 
0000034119 00000 n 
0000034246 00000 n 
0000034349 00000 n 
0000034477 00000 n 
0000034513 00000 n 
0000034639 00000 n 
0000034675 00000 n 
0000034803 00000 n 
0000034839 00000 n 
0000034967 00000 n 
0000035003 00000 n 
0000035131 00000 n 
0000035167 00000 n 
0000035295 00000 n 
0000035331 00000 n 
0000035459 00000 n 
0000035495 00000 n 
0000035623 00000 n 
0000035659 00000 n 
0000035787 00000 n 
0000035823 00000 n 
0000035951 00000 n 
0000035987 00000 n 
0000036113 00000 n 
0000036149 00000 n 
0000036277 00000 n 
0000036313 00000 n 
0000036441 00000 n 
0000036477 00000 n 
0000036605 00000 n 
0000036641 00000 n 
0000036769 00000 n 
0000036805 00000 n 
0000036931 00000 n 
0000036967 00000 n 
0000037095 00000 n 
0000037131 00000 n 
0000037259 00000 n 
0000037295 00000 n 
0000037423 00000 n 
0000037459 00000 n 
0000037586 00000 n 
0000037730 00000 n 
0000037856 00000 n 
0000037956 00000 n 
0000038083 00000 n 
0000038186 00000 n 
0000038314 00000 n 
0000038350 00000 n 
0000038476 00000 n 
0000038512 00000 n 
0000038640 00000 n 
0000038676 00000 n 
0000038804 00000 n 
0000038840 00000 n 
0000038968 00000 n 
0000039004 00000 n 
0000039132 00000 n 
0000039168 00000 n 
0000039296 00000 n 
0000039332 00000 n 
0000039460 00000 n 
0000039496 00000 n 
0000039624 00000 n 
0000039660 00000 n 
0000039788 00000 n 
0000039824 00000 n 
0000039950 00000 n 
0000039986 00000 n 
0000040114 00000 n 
0000040150 00000 n 
0000040278 00000 n 
0000040314 00000 n 
0000040442 00000 n 
0000040478 00000 n 
0000040606 00000 n 
0000040642 00000 n 
0000040768 00000 n 
0000040804 00000 n 
0000040932 00000 n 
0000040968 00000 n 
0000041096 00000 n 
0000041132 00000 n 
0000041262 00000 n 
0000041299 00000 n 
0000041428 00000 n 
0000041573 00000 n 
0000041701 00000 n 
0000041802 00000 n 
0000041931 00000 n 
0000042035 00000 n 
0000042165 00000 n 
0000042202 00000 n 
0000042330 00000 n 
0000042367 00000 n 
0000042497 00000 n 
0000042534 00000 n 
0000042664 00000 n 
0000042701 00000 n 
0000042831 00000 n 
0000042868 00000 n 
0000042998 00000 n 
0000043035 00000 n 
0000043165 00000 n 
0000043202 00000 n 
0000043332 00000 n 
0000043369 00000 n 
0000043499 00000 n 
0000043536 00000 n 
0000043666 00000 n 
0000043703 00000 n 
0000043831 00000 n 
0000043868 00000 n 
0000043998 00000 n 
0000044035 00000 n 
0000044165 00000 n 
0000044202 00000 n 
0000044332 00000 n 
0000044369 00000 n 
0000044499 00000 n 
0000044536 00000 n 
0000044664 00000 n 
0000044701 00000 n 
0000044831 00000 n 
0000044868 00000 n 
0000044998 00000 n 
0000045035 00000 n 
0000045165 00000 n 
0000045202 00000 n 
0000045473 00000 n 
0000067454 00000 n 
0000067571 00000 n 
0000067701 00000 n 
0000067738 00000 n 
0000067868 00000 n 
0000067905 00000 n 
0000068033 00000 n 
0000068070 00000 n 
0000068200 00000 n 
0000068237 00000 n 
0000068367 00000 n 
0000068404 00000 n 
0000068533 00000 n 
0000068570 00000 n 
0000068700 00000 n 
0000068737 00000 n 
0000068865 00000 n 
0000068976 00000 n 
0000075823 00000 n 
0000075953 00000 n 
0000075990 00000 n 
0000076120 00000 n 
0000076157 00000 n 
0000076285 00000 n 
0000076322 00000 n 
0000076452 00000 n 
0000076489 00000 n 
0000076619 00000 n 
0000076656 00000 n 
0000076785 00000 n 
0000076822 00000 n 
0000076952 00000 n 
0000076989 00000 n 
0000077117 00000 n 
0000077228 00000 n 
0000077358 00000 n 
0000077395 00000 n 
0000077525 00000 n 
0000077562 00000 n 
0000077690 00000 n 
0000077727 00000 n 
0000077857 00000 n 
0000077894 00000 n 
0000078024 00000 n 
0000078061 00000 n 
0000078190 00000 n 
0000078227 00000 n 
0000078357 00000 n 
0000078394 00000 n 
0000078522 00000 n 
0000078633 00000 n 
0000078832 00000 n 
0000097101 00000 n 
0000097231 00000 n 
0000097268 00000 n 
0000097398 00000 n 
0000097435 00000 n 
0000097565 00000 n 
0000097602 00000 n 
0000097731 00000 n 
0000097842 00000 n 
0000107844 00000 n 
0000107972 00000 n 
0000108083 00000 n 
0000116904 00000 n 
0000117034 00000 n 
0000117071 00000 n 
0000117201 00000 n 
0000117238 00000 n 
0000117368 00000 n 
0000117405 00000 n 
0000117534 00000 n 
0000117645 00000 n 
0000117773 00000 n 
0000117884 00000 n 
0000118014 00000 n 
0000118051 00000 n 
0000118181 00000 n 
0000118218 00000 n 
0000118348 00000 n 
0000118385 00000 n 
0000118514 00000 n 
0000118625 00000 n 
0000118753 00000 n 
0000118864 00000 n 
0000119111 00000 n 
0000140800 00000 n 
0000140930 00000 n 
0000140967 00000 n 
0000141097 00000 n 
0000141134 00000 n 
0000141264 00000 n 
0000141301 00000 n 
0000141431 00000 n 
0000141468 00000 n 
0000141598 00000 n 
0000141635 00000 n 
0000141764 00000 n 
0000141801 00000 n 
0000141931 00000 n 
0000141968 00000 n 
0000142098 00000 n 
0000142135 00000 n 
0000142265 00000 n 
0000142302 00000 n 
0000142432 00000 n 
0000142469 00000 n 
0000142599 00000 n 
0000142636 00000 n 
0000142766 00000 n 
0000142803 00000 n 
0000142932 00000 n 
0000142969 00000 n 
0000143099 00000 n 
0000143136 00000 n 
0000143266 00000 n 
0000143303 00000 n 
0000143433 00000 n 
0000143470 00000 n 
0000143600 00000 n 
0000143637 00000 n 
0000143767 00000 n 
0000143804 00000 n 
0000143934 00000 n 
0000143971 00000 n 
0000144100 00000 n 
0000144137 00000 n 
0000144267 00000 n 
0000144304 00000 n 
0000144371 00000 n 
0000166474 00000 n 
0000166541 00000 n 
trailer
<<
/Size 280
/Root 1 0 R
/Info 5 0 R
>>
startxref
176334
%%EOF