Fig 6. OCIAD1 is not involved in the HCV life cycle in vitro.

(A) Huh-7.5 cells were transfected with OCIAD1 siRNAs (#1, #2) or non-targeted control siRNA (nt) 24 h prior to transient transduction with recombinant retroviruses (multiplicity of infection [MOI] = 10) expressing either wild-type OCIAD1 (OCIAD1), uncleavable mutant C38A (C38A), truncated mutant N39 (N39) or the green fluorescent protein (GFP). Twenty-four h post-transduction, the cells were inoculated with cell culture-derived HCV (Jc1) at a MOI of 0.5 for 4 h. Cells were harvested 72 h post-infection and protein lysates were separated by 12% SDS-PAGE, followed by immunoblot with a polyclonal antibody against OCIAD1 or monoclonal antibody 9E10 against HCV NS5A. (B) Extra- and intracellular infectivities were determined 72 h post-infection by 50% tissue culture infective dose (TCID₅₀) determination. Each histogram represents the mean ± SEM of two independent experiments performed in triplicate.