FIG 6.

Knockdown of VPS4A decreases the release of infectious HCV particles. (A) Huh-7.5 cells were transfected with either VPS4A siRNA (80 nM) or VPS4B siRNA (40 nM). At 24 h posttransfection, cells were infected with HCV J6/JFH1 at an MOI of 2. The cell lysate and the culture supernatants were harvested at 6 dpi. The cell lysates were analyzed by immunoblotting with the indicated antibodies. The level of GAPDH served as a loading control. (B and C) Extracellular virus infectivity (B) and intracellular virus infectivity (C) were measured by a focus-forming assay. (D) Intracellular HCV RNA was quantitated by RT-qPCR. Amounts of intracellular HCV RNA were normalized to amounts of GAPDH mRNA. The value for the control cells was arbitrarily expressed as 1.0. Data represent means ± SEM of data from three independent experiments, *, P < 0.05, compared with the control.