Immortalized human myoblast LHCN-M2 cell lines and FSHD clonal cell lines were cultured in proliferation media [F10 (HyClone) supplemented with 20% fetal bovine serum (FBS) (PeakSerum, Ps-FB3, lot 293Q16), 1× 2-mercaptoethanol (Gibco), 10−9 M dexamethasone (Sigma-Aldrich), basic fibroblast growth factor (10 ng/ml; PeproTech), GlutaMAX (Gibco), and penicillin/streptomycin (Gibco)] at 37°C in a 5% CO2 atmosphere. Myogenic differentiation was initiated on confluent cell cultures with differentiating media [Dulbecco’s modified Eagle’s medium (DMEM)/F12; Corning Cellgro], supplemented with 1× Insulin-Transferrin-Selenium (Gibco), 1× non-essential amino acids (Gibco), GlutaMAX, and penicillin/streptomycin. 293T, A204-iDUX4, RH30-iDUX4, and RD-iDUX4 cells were cultured in DMEM/F12 supplemented with 10% FBS and penicillin/streptomycin. iC2C12-DUX4 and iC2C12-mDUX cells were cultured in DMEM (HyClone) and 20% FBS.

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