Skip to main content
. 2015 Feb 4;89(8):4387–4404. doi: 10.1128/JVI.02705-14

FIG 1.

FIG 1

AAK1 and GAK are essential for HCV entry. Huh-7.5 cells were transfected with siRNAs targeting either AAK1 (1 or 2), GAK, or PAK1 or with a nontargeting (NT) sequence. (A) Cells were lysed at 48 h posttransfection, and the indicated protein levels were measured by quantitative Western analysis. Representative membranes are shown. Numbers represent relative ratios of AAK1, GAK, or PAK1 to actin protein normalized to the NT control level from at least two independent measurements. (B) Cellular viability was measured in these cells by alamarBlue-based assays. Data are relative fluorescence values normalized to the NT control. (C) AAK1-, GAK-, or PAK1-depleted cells and NT controls were infected with HCV pseudoparticles (HCVpp) expressing a luciferase reporter gene for 4 h. Luciferase measurements were taken at 48 h postinfection. RLU, relative light units. (D) The indicated depleted cells and NT controls were infected with cell culture-grown HCV (HCVcc) expressing a luciferase reporter gene for 4 h. Luciferase measurements were taken at the indicated time points postinfection. (E) Huh-7.5 cells depleted of AAK1 or GAK and the NT controls were electroporated with in vitro-transcribed HCV RNA encoding a luciferase reporter. Luciferase measurements were taken at 6 and 72 h postelectroporation as a measurement of HCV RNA replication. Data represent means and standard deviations (error bars) from three experiments in triplicates. *, P < 0.05; **, P < 0.01; ***, P < 0.001.