Figure 4.

Impact of RNF20 knockdown on in vitro proliferation, migration, and invasion of lung cancer cell lines. (a) Western blot analysis of cell lysates of A549 and H460 cells transfected with either control or RNF20 siRNA and harvested at 72 h post-transfection. Actin served as loading control, total and phosphorylated Erk/Akt (T/P-Erk/Akt), p53, p21 E-Cadherin, and Vimentin were detected. The p53 blots were from separately developed blots for each cell, and the below Actin is its loading control. (b) Histogram of proliferation shows RNF20 knockdown increased the in vitro proliferation of both A549 and H460 cells over 72h (*P < 0.05, compared to control siRNA). (c) Representative photomicrographs of the migration chamber at the times indicated in control or RNF20 siRNA-transfected cells is shown on the left, with quantitative analysis of migrated cells shown on the right (N = 3 replicates per cell type). Error bars show SD (*P < 0.05, compared with control siRNA). (d) Lower surface of Matrigel transwell membranes seeded with cancer cells previously transfected with indicated siRNAs, 24 h after incubation (left), and corresponding quantitative analysis of invading cells (right). Data are shown as mean values graphed for indicated cells on the right (N = 3 replicates per cell type). Error bars show SD (*P < 0.05, ^**P < 0.01, compared with control siRNA).