FIG. 3.

Ago2 depletion also attenuates replication from a full-length JFH-1 genomic RNA. (A) Schematic representation of the full-length HCV JFH-1 RNA. RLuc, Renilla luciferase. (B) Huh 7.5 cells were electroporated with the indicated siRNAs. Three days later, the cells were reelectroporated with J6/JFH-1 Rluc RNA plus siRNA, and 2 days later, they were assayed for luciferase expression. The luciferase expression levels are presented relative to that seen in the sample treated with J6/JFH-1 Rluc and miControl. (C) Samples from the experiment in panel B prepared 2 days after the second electroporation were subjected to Northern blot analysis for HCV RNA using GAPDH RNA as a loading control (top) and by immunoblot analysis of human Ago2 protein with β-actin as a loading control (bottom). (D) Relative cell numbers in each sample from the experiment in panel B as measured by Wst-1 assay. (E) miR-122 levels 2 days after the second electroporation were analyzed in cells from panel B by real-time reverse transcriptase PCR. The data are shown as averages from four independent experiments; the error bars represent standard deviations.