Transcriptome analysis of Endothelin-3 or Neurotensin treated MuSCs

GPCRs have emerged as crucial regulators of muscle stem cell (MuSC) quiescence, yet the specific repertoire of GPCRs in quiescent MuSCs and the underlying regulatory network remain largely elusive. By employing pharmacological and inducible-genetic methods, we have identified two niche-derived GPCR ligands, endothelin-3 (ET-3) and neurotensin (NT), which bind to EDNRB and NTSR2 receptors, respectively, reinforcing the maintenance of MuSC quiescence. To comprehensively unravel the molecular mechanisms underlying the inhibitory effects of ET-3 and NT on MuSC activation, we conducted RNA sequencing (RNA-seq) analysis on FACS-purified MuSCs that underwent a 5-day in vitro treatment with ET-3 or NT After 5-day in vitro treatment with ET-3 or NT, mRNA profiles of the Ctrl (DMSO)or treated MuSCs were generated by deep sequencing, in triplicate, using NextSeq500 instrument (Illumina) using v2 chemistry with 1x75bp single end setup.