RPS3a Over-Expressed in HBV-Associated Hepatocellular Carcinoma Enhances the HBx-Induced NF-κB Signaling via Its Novel Chaperoning Function
Figure 5
Knock-down of RPS3a abolishes RPS3a-mediated NF-κB enhancement by HBx.
(A) A schematic representation of siRNA design on RPS3a gene. (B) The effect of three siRNA candidates on RPS3a, HBx and endogenous p65 expression. Cells were co-transfected with pcD-RPS3a-flag (or pcD-HBx-HA) and each RPS3a siRNA candidate (siRNA1, 2, 3), and harvested after 72 hr post-transfection. siRNAs against lamin, GFP, luciferase (GL3) were used as control siRNAs. Expression levels of proteins were detected by indicated antibodies. (C) Effect of RPS3a knock-down on NF-κB activity. The plasmids of pcD-RPS3a (0.4 µg), pEG-HBx (0.4 µg), and pNF-κB-Luc (0.25 µg) were co-transfected with the RPS3a siRNA1 or siRNA2. Lamin siRNA was used as a control siRNA. (D) Effect of endogenous RPS3a on HBx-mediated NF-κB activation. NF-κB activity was determined at 30 hr post co-transfection of pNF-κB-Luc and indicated plasmids with/without siRNAs in Huh7 cells. Note that knock-down of the endogenous RPS3a lowered the HBx-induced NF-κB activity to the background level.