RNA Interference and Single Particle Tracking Analysis of Hepatitis C Virus Endocytosis
Figure 5
HCV entry requires an intact actin cytoskeleton.
A. Huh-7.5 cells were incubated with indicated concentrations of cytochalasin D for one hour and infected with HCV pseudoparticles (HCVpp) expressing the luciferase reporter gene for four hours. Luciferase measurements were taken at 48 hours post infection. Plotted are relative luciferase values normalized to DMSO vehicle control. Cell viability was unaffected (data not shown). Asterisk indicates p<0.05, indicating significance. B. Huh-7.5 cells transiently expressing GFP-actin (green) were infected with DiD-HCV on ice for 1 hour then shifted to 37°C upon imaging. Scale bar = 10 µm. C. Montage of alternating time-lapse exposures of Cy5 (DiD-HCV) and GFP (actin) from inset in (B). Height of montage = 11.36 µm. Cells were treated with 2 µM cytochalasin D to depolymerize actin filaments at time indicated (arrow). Indicated times are in minutes: seconds. D. Time-lapse montage of (C), Cy5 (DiD-HCV) only. Experiment is representative of multiple replicates.