S1810

MCF-7 and MDA-MB-453 cell lines were obtained from ATCC (HTB-130 and HTB-22) and grown in DMEM 1 g/L glucose (Dominique Dutscher, L0066) containing fetal calf serum (5 %) (Dominique Dutscher, S1810), penicillin (50 U/ml) (Dominique Dutscher, L0018), streptomycin (50 μg/ml), and amphotericin B (1.25 μg/ml) (PAA, P11-001) at 37 °C in 5 % CO₂, and routinely used at 70–80 % confluence. When indicated, cells were exposed to 2 μM 5-aza-CdR (A3656, Sigma-Aldrich) for 48 h and 400 nM TSA (T8552, Sigma-Aldrich) for 16 h.

The A549 (NSCLC) cell line was obtained from Dr. Christophe Borg (INSERM UMR1098, Besançon, France) and the SiHa cells were obtained from ATCC (HTB-35™). A549 cells were grown in Dulbecco’s minimum essential medium (DMEM) 1 g/L glucose (L0066, Dominique Dutscher, Brumath, France) and SiHa cells were cultured in RPMI1640 medium (Dutscher, L0498). Both media were supplemented with fetal bovine serum (10%) (S1810, Dominique Dutscher). Cells were cultured at 37 °C in 5% CO₂ and routinely used at 70–80% confluence.
Transfections of the plasmids were performed using the Lipofectamine 2000 reagent (11668030, ThermoFisher) following the supplier’s recommendations. Briefly, cells were seeded in 6-well plates and transfected for 24 h with 1 µg of plasmid and 2 µL of Lipofectamine 2000 reagent and harvested 24 h after the end of the transfection. Transfections of the siRNA were conducted using the same protocol with 20 pmol of siRNA and 3 µL of Lipofectamine 200 reagent.

Human HeLa cells (CCL-2; ATCC) were obtained from the IGBMC cell culture facility and cultured in DMEM (4.5 g L^–1 glucose) supplemented with 10% fetal calf serum (Dutscher, S1810), 100 U ml penicillin and 100 μg ml^–1 streptomycin (Invitrogen, 15140-130). E14 mESCs (ES Parental cell line E14Tg2a.4, Mutant Mouse Resource and Research Center) were obtained from the IGBMC cell culture facility and cultured on gelatinized plates in feeder-free conditions in KnockOut DMEM (Gibco) supplemented with 20 mM l-glutamine, penicillin–streptomycin, 100 µM non-essential amino acids, 100 µM β-mercaptoethanol, N-2 supplement, B-27 supplement, 1000 U ml^–1 LIF (Millipore), 15% ESQ FBS (Gibco) and 2i (3 µM CHIR99021, 1 µM PD0325901, Axon MedChem). Cells were grown at 37 °C in a humidified, 5% CO₂ incubator.