FIGURE 6.

Immunofluorescence assay to examine CINP relocalization due to NS5B. A, GFP-CINP was co-transfected with equal amount of plasmids expressing myc-NS5B, myc-ΔNS5B, or empty vectors into Huh7 cells. Two days after transfection, the cells were fixed. Immunofluorescence staining was performed using the anti-myc antibody as a primary antibody. The cells were visualized by confocal laser-scanning microscopy. Green fluorescence indicates GFP-CINP, whereas red (Cy3) indicates NS5B or ΔNS5B. Yellow (merged) indicates colocalization. B, Huh7.5 cells, HCV subgenomic replicon cells, and JFH-1-preinfected Huh7.5 cells were transfected with GFP or GFP-CINP. Forty-eight hours later, cells were subjected to immunofluorescence staining as in A with the anti-NS5A antibody used as primary antibody. Green fluorescence indicates GFP or GFP-CINP, and red (Cy3) indicates endogenous NS5A. Yellow (merged) indicates colocalization.