siRNA Knockdown of ZNF703 Impacts Cell Proliferation

Cell lines were transfected with short-interfering RNA (siRNAs, 30 nM final concentration) in 6-well plates with RNAiMAX (Invitrogen) according to the manufacturer’s instructions and harvested 48 hours after transfection, which could be cultured to enter following experiments. Target sequences for the siRNA of ZNF703: sense strand-5’ CCACACACUUUGGGCCUAA dTdT 3’; antisense-strand-3’ dTdT GGUGUGUGAAACCCGGAUU 5’. Non-targeting control siRNA was designed and synthesized by Guangzhou RuiBoBio (Guangzhou, China). Proliferation assay and colony-forming assay were performed as previously described [22 (link)]. Cell proliferation was measured by sulforhodamine B (SRB) (Sigma) assay. Relative growth was calculated as the value relative to controlled cells. In colony-forming assay, cells were seeded into 6-well plates (1000 cells per well). After several proper days, colonies were fixed in 10% acetic acid, 10% methanol and 80% ddH2O, and then stained with crystal violet (0.5% w/v).