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Fig. 8 | Skeletal Muscle

Fig. 8

From: Neuromuscular electrical stimulation promotes development in mice of mature human muscle from immortalized human myoblasts

Fig. 8

Centrally located myonuclei of human and murine origin in xenografts. Unfixed cross sections were immunolabeled with antibodies to human β-spectrin (green) and human lamin A/C (red) and counterstained with DAPI (blue) to label all nuclei. Human myonuclei, positive for DAPI and human lamin A/C, and murine myonuclei, positive for DAPI but not for human lamin A/C, were quantitated in human myofibers and identified by the presence of β-spectrin at the sarcolemma. Images of human myofibers in group 2 (a, b) and group 3 (c, d) show labeling for spectrin and lamin A/C (a, c) and for both proteins and DAPI (b, d). a, b Arrows point to myofibers with nuclei labeled with antibodies to human lamin A/C. Arrowheads point to nuclei labeled with antibodies to human lamin A/C lying outside of muscle fibers. c, d Arrow points to a human myofiber with a nucleus that is unlabeled by antibodies to human lamin A/C. Arrowheads point to nuclei that lie outside of muscle fibers and that do not label with antibodies to human lamin A/C. e High magnification examples of human fibers with a myonucleus that labels with anti-human lamin A/C (E1), a nucleus lying outside a human fiber that labels with anti-human lamin A/C (E2), a human fiber with a myonucleus that fails to label for human lamin A/C (E3), and a nucleus that fails to label for human lamin A/C lying outside a human fiber (E4). f Percent of centrally located human and murine myonuclei detected in human muscle fibers in groups 2 and 3 at 4–5-weeks post-transplantation. Of the human fibers examined, <20 % was centrally nucleated (group 2 = 36/205, or 17.5 %; group 3 = 69/505 or 13.7 %). Only 2 myonuclei out of 205 (1 %) in group 2 and 7 myonuclei out of 505 (1.4 %) in group 3 failed to label with antibodies to human lamin A/C. Scale bars = 10 μm

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