Figure 6

Diprotin A enhances proliferation of donor cells in vivo. Mouse muscle injected with 1 × 10⁴ canine muscle-derived mononuclear cells alone, or cells treated with 5 mM diprotin A, were harvested weekly for 4 weeks after injection (A, B, C), or at 3, 5, or 7 weeks after injection (D, E, F) and cryosections probed with anti-dystrophin (MANDYS107), anti-lamin A/C, and/or Pax7, and fluorescently labeled secondary antibody. The number of fibers expressing canine dystrophin (A, D), and the number of nuclei expressing canine lamin A/C (B, E), and the number of nuclei co-expressing Pax7 and canine lamin A/C (C, F) per cross-section were determined, and the points represent the average of the averages ± SD, where the average was calculated from three cryosections per mouse, and the average of the averages was calculated from three mice per condition. (G, H) Mice injected with 1 × 10⁴ canine muscle-derived mononuclear cells alone, or cells treated with 5 mM diprotin A, were given EdU daily for 1 week during weeks 1, 2, 3, or 4 after injection. Injected muscle was harvested after the last dose of EdU, and cryosections stained for canine lamin A/C, or Pax7 and canine lamin A/C, and EdU. The percentage of canine lamin A/C-positive nuclei that incorporated EdU (G) and the percentage of canine lamin A/C:Pax7 double positive nuclei that incorporated EdU (H) per cross-section were determined. The points represent the average of the averages ± SD, where the average was calculated from three cryosections per mouse, and the average of the averages was calculated from three mice per condition. The P value is the result of a Student's t-test.