Fig 6.

Rab1 activity is essential for LD biogenesis and HCV replication. (A) The dominant negative Rab1 N121I mutant blocks LD biogenesis. Huh7 cells were transfected with wild type (WT; top) or dominant negative (N121I; bottom) GFP-Rab1 (green). At 24 h posttransfection, images of living cells were captured after labeling for 20 min with 20 μg/ml Bodipy (red). (B) Huh7.5 cells were transfected with the J6/JFH genome by electroporation of 10 μg in vitro-transcribed J6/JFH RNA. After 24 h, the infected cells were transfected with wild-type (top) or dominant negative (N121I, bottom) GFP-Rab1 (green). At 24 h posttransfection, cells were fixed, permeabilized, and analyzed by immunofluorescence using anti-NS5A-specific antibody (middle and red). The contour of the Rab-GFP expressing cells is outlined in yellow. Bars = 10 μm.