FIG. 1.
Effect of siRNAVP7 and siRNANSP4 on the synthesis of viral proteins. MA104 cells in 48-well plates were transfected with the indicated siRNA, and after 8 h, the Lipofectamine-siRNA mixture was removed and replaced by MEM. Forty-eight hours posttransfection, the cells were infected with rotavirus RRV at an MOI of 3, and 12 h postinfection, the cells were harvested and processed for SDS-PAGE. (A) Fluorography of cells transfected with the indicated siRNAs in which the cells were labeled with 50 μCi of Easy Tag EXPRESS-35S/ml at 4 hpi and harvested 12 hpi. (B) A lysate of cells transfected with the indicated siRNAs was harvested at 12 hpi and subjected to SDS-PAGE. The proteins were transferred to nitrocellulose membrane and immunostained with a rabbit polyclonal antibody to rotavirus NSP5. (C) Immunoblot analysis of lysates of cells transfected with the indicated siRNAs and harvested at 12 hpi. The proteins were incubated with a rabbit anti-rotavirus polyclonal antiserum. The relative positions of the viral proteins are indicated in each gel. Ctrol, control.