Fig. 6.

Validation of the silencing efficiency of selected shRNA targeting HCV cofactors and their effect on viral replication. A, the gene silencing effect was evaluated for each cofactor via Western blot detection. A decrease of the target protein was observed with the gene-specific shRNA, illustrating the efficacy of gene silencing. Actin immunoblot was performed as a loading control. B, C, the gene silencing effects of shRNA-expressing lentiviruses for each cofactor were determined on HCV replication-dependent reporter activities (MOI of 10) at day 3 post-transduction with HCV replicon-containing cells (B) and in the infection assay (C). The cellular viability was assessed with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxicity assay performed in parallel.