G 6264

R1 mESCs, ZHBTc4 (mESC lines) (Niwa et al., 2000 (link)) and pre-iPS cells were cultured on gelatin-coated tissue culture plates in knockout DMEM (Thermo Fisher, 10829018) supplemented with 15% fetal calf serum (Lonsera, S711-004S/NN02953), 1× nonessential amino acids (Gibco, 11140050), 2 mmol/L L-glutamine (Gibco, 35050061), 1% (v/v) nucleoside mix (Sigma-Aldrich, A-4036, T-1895, C-4654, G-6264, U-3003), 0.1 mmol/L β-mercaptoethanol (Sigma-Aldrich, M6250), 1,000 U/mL recombinant leukemia inhibitory factor (LIF) (Millipore, ESG1107).

Mouse PSCs were cultured on Mitomycin C (M4287, Sigma-Aldrich)-inactivated mouse embryonic fibroblast (MEF) feeders in ESC medium consisting of Dulbecco’s modified Eagle’s medium with high glucose, pyruvate (DMEM, 11995065, Gibco), 15% fetal bovine serum (FBS, TMS-013-BKR, Millipore), 1,000 U/mL mLIF (ESG1107, Millipore), 1% Penicillin-Streptomycin (15140-122, Gibco), 2 mM L-glutamine (A29168-01, Gibco), MEM NEAA (11140-050, Gibco), 1 mM sodium pyruvate (11360-070, Gibco) and 0.1 mM 2-mercaptoethanol (ES-007-E, Millipore). The cells were maintained in a 37 °C humid incubator with 5% CO₂, and the culture medium was changed every two days. Retinoic acid (RA, R2625, Sigma-Aldrich), 2-deoxyglucose (2DG, D6134, Sigma-Aldrich), 6-aminonicotinamide (6AN, A68203, Sigma-Aldrich) and guanosine (G6264, Sigma-Aldrich) were used in the experiments.