Integrin α7 allophycocyanin

Primary MuSCs were isolated as previously described.²⁴ Briefly, TA muscles from 3‐month‐old mice were dissected and dissociated with collagenase (Roche, Indianapolis, IN, USA). The cells were negatively selected by biotinylated CD45, CD11, CD31 and Sca1 antibodies. The muscle cells in the flowthrough were subjected to CD34‐FITC (BD biosciences) and integrin α₇‐allophycocyanin (R&D systems, Minneapolis, MN, USA) staining. The viable PI⁻CD34⁺integrin‐α₇⁺ MuSCs were collected by FACS sorting (Influx, BD biosciences, Franklin Lake, NJ, USA). MuSCs were cultured on collagen coated dishes in F10 medium containing 10% foetal bovine serum (FBS), 5 ng/mL IL‐1α, 5 ng/mL IL‐13, 10 ng/mL IFN‐γ, and 10 ng/mL TNF‐α (R&D Systems), and 2.5 ng/mL FGF (Invitrogen, Red Wood City, CA, USA) as described previously.²⁴ MuSCs were differentiated in differentiation medium [DMEM medium (invitrogen)] containing 2% horse serum (HyClone, Malborough, MA, USA) for 3 days. C2C12 cells (ATCC) were cultured in DMEM medium (Invitrogen) containing 10% FBS (HyClone) and differentiated in DMEM medium (Invitrogen) containing 2% horse serum (HyClone) for 3 days. The differentiated myotubes were further isolated by pre‐attaching to plates for three times.

Primary MuSCs were isolated as previously described. Briefly, TA muscles from 3-months-old C57BL/6 mice were dissected and dissociated with collagenase (Roche, Indianapolis, IN, USA). The muscle cells in the flowthrough were subjected to CD34-FITC (BD biosciences) and integrin α₇-allophycocyanin (R&D systems, Minneapolis, MN, USA) staining. The viable PI⁻CD34⁺integrin-α₇⁺ MuSCs were collected by FACS sorting (Influx, BD biosciences, Franklin Lake, NJ, USA). MuSCs were cultured on collagen coated dishes in F10 medium containing 10% FBS, 5 ng/ml IL-1α, 5 ng/ml IL-13, 10 ng/ml IFN-γ, and 10 ng/ml TNF-α (R&D Systems), and 2.5 ng/ml FGF (Invitrogen, Red Wood City, CA, USA) as described previously. MuSCs were differentiated in differentiation medium (DMEM medium (invitrogen) containing 2% horse serum (HyClone, Malborough, MA, USA) for 3 days. C2C12 cells (ATCC) were cultured in DMEM medium (Invitrogen) containing 10% FBS (HyClone), and differentiated in DMEM medium (Invitrogen) containing 2% horse serum (HyClone) for 3 days. The differentiated myotubes were further isolated by pre-attaching to plates for 3 times.