FIG. 7.
siRNA knockdown of ADAR1 stimulates replicon expression. (A) Huh.BB7 cells were plated at 1 × 106 and transfected once or twice with siRNA directed towards ADAR1 or with siRNA directed against ADAR2. The histogram shows Taqman analysis of HCV RNA from equal numbers of cells. The number of transfections is shown below the bars in the histogram. (Β) Twenty-five micrograms of cytoplasmic lysates shown in panel A. siRNA experiments were repeated three times with similar results. (C) Huh.BB7 cells were transfected with ADAR1-specific siRNA and treated with IFN-α (10 or 1,000 IU/ml) at 4 days posttransfection. Cells were harvested after 18 h of IFN-α treatment. RNA was isolated and monitored by Taqman analysis (16). (D) Cytoplasmic extracts were analyzed for protein expression as described in Materials and Methods. The migration positions of molecular mass markers are shown to the left of the blots.