Antiviral activity of Pol III promoter-driven siRNAs and tsiRNAs in HCV replicon cells, FK-R2AN. (A) tsiRNA-mediated inhibition of HCV replication in FK/R2AN cells. HCV replicon cells expressing the Renilla luciferase reporter were transfected with 1 μg of the siRNA- or tsiRNA-expression pGD plasmids indicated. On day 2, cell lysates were harvested for the Renilla luciferase assay. Reporter protein expression levels of cells transfected with (left) the control siRNA (siC) or (right) tsiRNA (tsiC) construct were set at 100% in each graph. Assays were performed in triplicate. (B) tsiRNA-mediated inhibition of HCV core gene expression. Immunoblotting was performed using a core-specific antibody and an antibody against β-actin (an internal loading control). (C) tsiRNA-mediated inhibition of HCV NS5A gene expression in FK/R2AN cells. Cells transfected with representative tsiE2N RNA-expressing pGD vectors indicated were immunostained with rabbit anti-NS5A polyclonal antibody and goat anti-rabbit IgG-Texas Red (red) on day 2.
