Figure 2

Effects of Plexin-B1 siRNA on SKOV3 cells. (A) Real-time PCR was performed to detect Plexin-B1 mRNA levels in non-transfected cells (Blank control) and cells at 24 h after a 5 h exposure to negative control siRNA or one of the three different Plexin-B1 siRNAs (Plexin-B1 siRNA1-3). The mean relative level ± SD of each group is shown. Plexin-B1 mRNA levels in three Plexin-B1 siRNA groups were significantly down-regulated (*P < 0.01), while the negative control siRNA did not cause an obvious change. The bar graph shows the results of three independent experiments. (B) Western blot analysis of lysates from untransfected (Blank control) SKOV3 cells and SKOV3 cells at 24 h after a 5-h exposure to one of the three different Plexin-B1 siRNAs (Plexin-B1 siRNA1-3) or to negative control siRNA. The cells were analyzed by immunoblotting with specific antibodies to Plexin-B1, p-AKT (s473), AKT and β-actin. Plexin-B1 expression was significantly down-regulated in each of the three Plexin-B1 siRNA groups relative to the blank control group or the negative control group. p-AKT (s473) expression sequentially decreased in the three Plexin-B1 siRNA groups, and AKT changed impalpably at the protein level. β-actin was the internal loading control. (C) The western blots were scanned and quantified. Data present densitometric analyses of Plexin-B1, p-AKT (s473) or AKT relative to β-actin for n = 5 independent experiments. * indicates P < 0.01 when compared to the blank control.