Fig. 6

PPP2R5D enhanced HCV replication. (A and B) siRNA silencing of PPP2R5D decreased HCV replication in replicon-harboring cells. HCV genotype 2a replicon JFH1-SGRep replicon expressing cells were generated and used for silencing experiment. siRNAs targeting different regions of PPP2R5D (#1, #2, and #3) were transfected into the replicon cells for 36 h. The expression of NS5B protein level was detected by western blot and normalized to GAPDH (panel A), and viral RNA levels were determined using qRT-PCR and relative to that in the control siRNA-treated cells (panel B). (C and D) Over-expression of PPP2R5D increased HCV replication in replicon cells. HA-PPP2R5D was transfected into HCV replicon cells for 36 h, HA-EGFP being a transfection control, and then HCV replication was evaluated for NS5B protein by western blot and normalized to GAPDH (panel C), and viral RNA levels were determined and relative to that of EGFP-transfected control cells (panel D). Images of western blot are representative of three independent experiments, while the data of viral RNAs was mean value with SEM of three independent experiments