Fig. 4: Aged MuSCs present decreased ciliation and Hedgehog signaling.

a Representative confocal images of uninjured/resting EDL myofibers of young (2 months) and aged (>24 months) C57BL/6 wild-type (wt) mice showing cilia immunostaining in Pax7⁺ MuSCs. Scale bars: 10 μm. DAPI, blue; PAX7, white; FOP, green; detyrosinated tubulin, red. b Percent of ciliated Pax7+ MuSCs quantified from isolated myofibers (n = 58 myofibers total per age group isolated from n = 4 young and n = 5 aged mice, average percent of ciliated MuSCs per mouse is shown). c Heat map of differentially expressed Hh pathway genes in young and aged MuSCs. d Expression of the negative regulator of Hh signaling, Ptch1, is increased in aged MuSCs, while the target gene Gli2 is decreased in aged MuSCs compared to young as determined by qPCR analysis (n = 4 mice per age group). e, f Expression of Ptch1 (e) and Gli2 transcription factor (f) after Tibialis anterior (TA) muscle injury (notexin) (n = 3–7 mice per timepoint and age group, individual data points in Supplementary Fig. 4c, d). *P < 0.05, **P < 0.01, ***P < 0.001 ****P < 0.0001. Mann–Whitney test (b, d). ANOVA test for group comparisons and significant difference for each timepoint relative to Day 0 by Fisher’s LSD test (e, f). Source data are provided as a Source Data file. Means+s.e.m.