Extended Data Fig. 5: Transcriptional effects of aging and exercise in quiescent MuSCs.
a, RT-qPCR in MuSCs from mice independent of those used in the RNA-Seq experiment. Ct values were normalized first to the mean of Gapdh, Hprt, and Actb1 and then to the mean Y(-Ex) level in each experiment, with Y(-Ex) shown as a dotted line at relative expression 1.0 for comparison (n=13 for O(-Ex), 14 for O(+Ex), and 13 for Y(-Ex) mice). b, GSEA results for the Hallmark gene sets in comparisons of RNA-Seq profiles for O(-Ex) vs. Y(-Ex), O(+Ex) vs. O(-Ex), and Y(+Ex) vs. Y(-Ex) MuSCs. Gene sets are in descending order based on the O(+Ex) vs. O(-Ex) NES. c, Enrichment plots for the INFLAMMATORY RESPONSE gene set. d, Single-cell RT-qPCR for Ccnd1 in freshly isolated MuSCs. For comparison, also shown are results for young MuSCs isolated three days after injury. The pairs on each chip were O(-Ex) vs. O(+Ex) and Y(-Ex) vs. Injured (n=24 cells from one mouse in each group). Data are summarized with mean and s.e.m. in a, box-and-whisker plots (bottom whisker, min; box bottom, 25th percentile; box middle, median; box top, 75th percentile; top whisker, max; “+”, mean) in d. NES, normalized enrichment score in b, c; ES, running enrichment score; S2N, GSEA Signal2Noise ranking metric in c. *P<0.05; two-tailed Welch’s t-test in a, d.
