Mouse myoblast C2C12 cells were obtained from American Type Culture Collection (ATCC, CRL-1772) and maintained in DMEM with 10% FBS in a 37°C and 5% CO2 incubator. This and other cell lines below were authenticated by each provider. To induce differentiation into myotubes, the cells at 90% confluency (day 0) were rinsed twice with phosphate buffered saline (PBS) before addition of 5% HS in DMEM. Medium was changed every two days.
Mouse embryonic stem cells CGR8 and ES-E14TG2a were purchased from European Collection of Cell Cultures (ECACC, 07032901) and ATCC (CRL-1821), respectively. They were cultured in Glasgow Minimum Essential Medium supplemented with 2 mM glutamine, 0.05 mM 2-mercaptoethanol, 1,000 units/ml leukemia inhibitory factor (MilliporeSigma, ESG1106), and 10% FBS in culture dishes coated with 0.2% gelatin in PBS for 15 min.
Mouse embryonic stem cells CGR8 and ES-E14TG2a were purchased from European Collection of Cell Cultures (ECACC, 07032901) and ATCC (CRL-1821), respectively. They were cultured in Glasgow Minimum Essential Medium supplemented with 2 mM glutamine, 0.05 mM 2-mercaptoethanol, 1,000 units/ml leukemia inhibitory factor (MilliporeSigma, ESG1106), and 10% FBS in culture dishes coated with 0.2% gelatin in PBS for 15 min.