Figure 5
Diprotin A enhances engraftment of donor cells. (A) 1 × 105 canine bone marrow cells (BMC), and 1 × 105 freshly isolated skeletal muscle-derived cells (SkMC) alone, or with 5 mM diprotin A (SkMC+dpA), were incubated with Gly-Pro-p-nitroaniline for 2 h. The nmol of p-nitroaniline (pNA) produced by peptidase cleavage was determined by measuring absorbance at 405 nm and converting the value to nmol of pNA using a standard curve. The bars represent the average nmol of pNA produced from three separate samples, and the P values are the result of a Student's t-test. (B, C, D) Cryosections from mouse muscle injected with 1 × 104 or 5 × 104 canine muscle-derived mononuclear cells alone or cells treated with 5 mM diprotin A were probed with anti-dystrophin (MANDYS107), anti-lamin A/C, or anti-lamin A/C and anti-Pax7, and fluorescently labeled secondary antibody. The number of fibers expressing canine dystrophin (B), the number of nuclei expressing canine lamin A/C (C), and the number of nuclei expressing canine lamin A/C and Pax7 (D) per cross-section were determined. The bars 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 values are the result of a Student's t-test. (E, F) Cryosections from mouse muscle injected with 1 × 104 freshly isolated mixed canine muscle-derived mononuclear cells alone, cells treated with 5 mM diprotin A, or cells treated with 5 mM diprotin A and α-CXCR4 antibody, were probed with anti-dystrophin (MANDYS107), or anti-lamin A/C, and fluorescently labeled secondary antibody. The number of fibers expressing canine dystrophin (E) and the number of nuclei expressing canine lamin A/C (F) per cross-section were determined. The bars 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 values are the result of a Student's t-test.