FIG. 6.

p62 knockdown results in reduced huTRIM5α-mediated restriction in HeLa cells. (A) HeLa cells were transfected with control or p62-specific siRNA, and the extent of knockdown was analyzed by Western blotting. (B) siRNA-transfected cells were plated in equivalent numbers and infected with B-tropic or N-tropic MLV overnight. Eighteen hours following infection, cells were harvested and viral DNA products were analyzed by quantitative PCR. Values were normalized to the amounts of β-actin DNA in identical samples. Error bars represent the standard deviations from triplicate samples. Values given above the columns represent the fold enhancement relative to the level for the untreated sample of that cell type. (C) Thirty-six to 48 h following infection, untreated (squares), control siRNA-transfected (circles), and p62 siRNA-transfected (triangles) cells were harvested and analyzed for GFP expression by flow cytometry. Ten thousand events/sample were analyzed. The percentage of GFP-positive cells is indicated for each sample. The dilution factor of virus used is given on the x axis. The data shown here are representative of three independent experiments.