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. 2010 May 11;5(5):e10575. doi: 10.1371/journal.pone.0010575

Figure 3. HCV induces IFN during the first 12 hrs of infection and inhibits it thereafter.

Figure 3

Huh7.25.CD81 and Huh7.5 cells were transfected with the pGL2-IFNβ-FLUC/pRL-TK-RLUC reporter plasmids together with plasmids expressing HA-TRIM25 (Huh7.25.CD81; A and B) or RIG-I (Huh7.5; C and D). 24 h post-transfection, the cells were infected with SeV (40 HAU/ml) or JFH1 (m.o.i = 0.2). A and C: 24 hrs post-transfection, the cells were infected with SeV (40 HAU/ml) or JFH1 (m.o.i = 0.2). At the times indicated, cell lysates were prepared and analysed for IFN induction as described in Materials and Methods. The graphs represent the levels of F-luc activity normalized to R-luc RNA expressed as IFN-β fold-induction over control cells that were simply transfected with pGL2-IFNβ-FLUC/pRL-TK-RLUC. Error bars represent the mean ± S.D. for triplicates. In addition, cell lysates from JFH1-infected cells were pooled and analysed for the presence of NS3 as a marker of HCV infection. B and D: 24 hrs post-transfection, Huh7.25.CD81 and Huh7.5 cells were infected with JFH1 (m.o.i = 0.2). At the times indicated, cells were processed for RNA extraction and HCV or IFNβ RNA were quantified by qRT-PCR respectively, and normalized against RNA from GAPDH. Error bars represent the mean ± S.D. for triplicates.