FIG 6.

Effects of TRAF6 on the NF-κB promoter and the expression of cytokines in HCV-infected cells. (A) Huh7 cells were transfected with the NF-κB-luciferase reporter and either the control siRNA or the TRAF6 siRNA for 24 h. The CMV-renilla luciferase reporter was also used for cotransfection to monitor transfection efficiencies. The cells were then infected with HCV for 24 h or 48 h and then lysed for measuring the luciferase activities using the Dual-Luciferase assay kit (Promega). The results represent the means plus SEM from three independent experiments. Huh7 cells transfected with the control siRNA or the TRAF6 siRNA were infected with HCV for 24 or 48 h. (B) Cells were then lysed for quantification of IL-6 or TNF-α (TNFa) mRNAs using real-time RT-PCR. The levels of IL-6 and TNF-α mRNAs of mock-infected cells transfected with the control siRNA were arbitrarily set at 1. (C) The incubation medium was collected and filtered at 24 and 48 h postinfection for analysis of IL-6 and TNF-α by ELISA.