Figure 4.

Effect of a recombinant adenovirus expressing shRNA on HCV replicon. (a) Huh7/pRep‐Feo cells were infected with AxshRNA‐HCV or shRNA‐Control at a multiplicity of infection (MOI) of 1. The cells were harvested, and internal luciferase activities were measured on day 0 though day 9 after adenovirus infection. Each assay was done in triplicate, and the value is displayed as a percentage of no treatment and as mean ± SD. An asterisk indicates a P‐value of less than 0.05. (b) Dimethylthiazol carboxymethoxyphenyl sulfophenyl tetrazolium (MTS) assay of Huh7/pRep‐Feo cells. Cells were infected with indicated recombinant adenoviruses at an MOI of 1. The assay was done at day 6 of infection. Error bars indicate mean + SD. (c) Northern blotting. The upper panel shows replicon RNA, and the lower panel shows beta‐actin mRNA. (d) Western blotting. Total cell lysates were separated on NuPAGE gel, blotted and incubated with monoclonal anti‐NS4A or anti‐NS5A antibodies. The membrane was re‐blotted with antibeta‐actin antibodies. NT, untreated Huh7/pRep‐Feo cells; Control, cells infected with AxshRNA‐Control; HCV, cells treated with AxshRNA‐HCV. In panels (b) and (c), cells were harvested on day 6 after adenovirus infection at an MOI of 1.