Fig. 2

Assaying satellite cell self-renewal and differentiation in vivo. a Representative FACS profile showing the percentage of ZsGreen+ cells that are lineage negative, double positive cells for the three most-commonly used antibodies to identify satellite cells (n=3). b Representative FACS profile for the isolation of purified satellite cells from transplanted muscle. Lineage-negative gating (against CD31 and CD45, both PE-Cy7 conjugated) is shown at left. Positive gating for VCAM-1 (PE), Itga7 (APC) is shown in the middle, and ZsGreen within the Lin– VCAM+Itga7+ population is shown at right. c Representative image showing Dystrophin (red) expression 4 weeks after transplant of 300 Pax7-ZsGreen cells. Scale bar is 100 μm. d FACS quantification of the number of ZsGreen+ cells after transplant with or without irradiation. Data shown are mean ± SE (n=12 transplanted TAs/group). Statistical comparisons were performed using two-tailed t tests. Note that irradiation significantly enhances contribution to the satellite cell pool. e Quantification of the total number of Dystrophin+ fibers 4 weeks after transplant in the presence/absence of irradiation. Shown are mean ± SE (n=12 transplanted TAs/group). Statistical comparisons were performed using two-tailed t tests. f Quantification of the number of ZsGreen+ cells 4 weeks after transplant of different numbers of ZsGreen+ cells. Shown are mean ± SE (n=6 transplanted TAs)