Figure 2.

Necroptosis and Apoptosis of MuSCs Ameliorate Muscle Function in Chronic Muscle Disease

(A) Schematic representation of the tamoxifen regimen.

(B) Immunofluorescence staining of TA muscle cross-sections from mdx and mdx/Ripk3^mKO mice 2 weeks after tamoxifen treatment using antibodies against PAX7 and cleaved CASP3 (n = 3 for each group). Scale bar. 25 μm.

(C) Quantification of apoptotic CASP3⁺/PAX7⁺MuSCs in TA muscle cross-sections.

(D) Quantification of total PAX7⁺ MuSCs in TA muscle cross-sections.

(E) Schematic representation of the tamoxifen regimen, z-VAD inhibitor treatment, muscle grip strength measurement, and time points of TA muscle collection.

(F–I) Immunofluorescence staining of TA muscle cross-sections from mdx and mdx/Ripk3^mKO mice for PAX7 and cleaved CASP3 (F) and RIPK3 (H) 2 weeks after tamoxifen treatment and DMSO or z-VAD inhibitor treatment (n = 3 or 4 for each group) and quantified in (G) and (I), respectively. Scale bar, 25 μm.

(J) H&E staining of TA muscles sections from mdx and mdx/Ripk3^mKO mice treated with DMSO or z-VAD inhibitor (n = 3 for each group). Scale bar, 100 μm.

(K) Quantification of muscle grip strength of mdx and mdx/Ripk3^mKO mice treated with DMSO or z-VAD inhibitor (n = 3 or 4 for each group).

Statistical analysis: ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.005, two-way ANOVA followed by Bonferroni post-test with alpha = 5%. All analyses indicated across the experiments were biological replicates unless otherwise stated.