GSM4654907: Prx1-Cre;Bap1f/f male#2; Mus musculus; RNA-Seq
Hindlimb muscles were mechanically dissected and dissociated in Dulbecco's modified Eagle's medium (DMEM) containing 10% horse serum (Hyclone, Logan, UT, USA), collagenase II (800 units/mL; Worthington, Lakewood, NJ, USA), and dispase (1.1 units/mL; StemCell Technologies, Vancouver, BC, Canada) at 37°C for 60 min. Digested suspensions were subsequently triturated and washed with DMEM to harvest mononuclear cells. Mononuclear cells were stained with Sca1-FITC (clone D7; BioLegend, San Diego, CA, USA), CD31-APC (clone MEC 13.3; BD Biosciences, Franklin Lakes, NJ, USA), CD45-APC (clone 30-F11; BD Biosciences), and Vcam1-Biotin (clone 429; BD Biosciences). Streptavidin-PE/Cy7 (BioLegend) was used as a secondary reagent. Stained cells were analysed and Lin–Vcam-Sca1+ cells were isolated using a FACS Aria III cell sorter (BD Biosciences). Total RNA was extracted from freshly isolated MuSCs using a TRIzol Reagent (Life Technologies, Carlsbad, CA, USA) RNA libraries were prepared for sequencing using standard Illumina protocols.