Synchronize Cancer Cell Cycle for Transfection

Prior to transfection, cells were starved for 6 h in order to achieve proper cell cycle synchronization. T98G cancer cells were transfected overnight with Dharmacon's chemically synthesized siRNA SMARTpools [human PC‐1, L‐007666‐00‐0005, ON‐TARGETplus Human PKD1 (5310) siRNA–SMARTpool, 5 nmol] and non‐targeting siRNA for control cells (D‐001210‐01‐05, siGENOME Non‐Targeting siRNA #1, 5 nmol), in dilution 1:20 in 1× siRNA buffer, using DharmaFECT 2 Transfection Reagent, 0.2 ml (Dharmacon) in dilution 1:50 in DMEM (Gibco, Thermo Fisher Scientific).18 (link), 19 (link) After 16 h, the medium was changed and the cells were treated with IgPC1 and/or HP and cultured for 24 and 48 h.