Cell line generation and cultivation for RanBPM study

Hela control shRNA and RanBPM shRNA stable cell lines (2–7 and 2–6) were described previously [29 (link),32 (link)] and were cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 0.35 mg/ml G418 (Geneticin, Bioshop Canada, Burlington, ON, Canada) at 37°C in 5% CO₂. 3T3 mouse embryonic fibroblasts (MEFs) were cultured in high-glucose DMEM supplemented with 10% FBS. Plasmid transfections were carried out with ExGen500 (Fermentas, Burlington, ON Canada), TurboFect Transfection Reagent (Thermo Fisher Scientific, Burlington, ON, Canada) or jetPRIME (Polypus Transfection) according to the manufacturer’s protocol. Leptomycin B (LMB, Bioshop Canada, Burlington ON, Canada) was added to the cells’ media at 20nM concentration for the times indicated in the figure legends. Nocodazole (Abcam) was added to the cell’s media at 10μM for 4 hours as previously described [33 (link),34 (link)].