FIG 7.

AAK1 and GAK are molecular targets underlying sunitinib and erlotinib antiviral effects. Huh-7.5 cells plated in six-well plates were transfected with plasmids encoding AAK1, GAK, or EGFR or with an empty plasmid. At 24 h posttransfection cells were trypsinized and plated in 6- and 96-well plates. (A) Western analysis was performed in cell lysates (obtained from the six-well plate) at 48 h posttransfection to assess the level of overexpression. Representative membranes immunoblotted (IB) with antibodies targeting the indicated proteins and actin are shown. (B) At 48 h posttransfection cells plated in 96-well plates were infected with HCVcc for 1 h on ice, followed by a temperature shift to 37°C and 4-h treatment with sunitinib (left) or erlotinib (right) at 0.5 μM and 1 μM or with a DMSO control. Medium was then replaced for removal of residual drugs, and unbound virus and luciferase signals were measured following a 20-h incubation. Data are relative luciferase values normalized to the values of the DMSO controls overexpressing the respective protein. Means and standard deviations (error bars) of results from three independent experiments are shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001.