Cells were pelleted at 300g for 5 minutes before washing with ice cold Hanks' Balanced Salt Solution without magnesium and calcium supplemented with 2% FBS and 1mM EDTA (HBSS+), pelleting again, and resuspending in 4ml of HBSS+. Samples were kept on ice and counterstained with SYTOX Blue before FACS isolation of MuSCs. Each sample was pooled from two littermate mice of the same genotype with 2 samples sequenced per genotype. A Sony MA900 equipped with a 100μm nozzle and 20 psi pressure was used for FACS isolation. Droplet-based scRNA-seq (Chromium, 10x Genomics) was applied to single-cell suspensions of tdTomato+ cells from 2 WT and 2 α-cdKO samples (2 mice pooled per sample); 3 × 10^4 cells were loaded per well for each of the 4 samples, and partitioning and library preparation were performed according to the manufacturer's protocol for the 3′ v.3 chemistry.