Figure 4. OSBP inhibition alters web morphology without blocking HCV-induced PI(4)P accumulation.
(A) Nonreplicative expression of the NS3-5B(NS5A/GFP) polyprotein in Huh7.5.1/T7 cells stably expressing T7 RNA polymerase and the indicated shRNAs. 48 hr post-transfection, cells were immunostained for PI(4)P (red) and GFP (green) with DAPI nuclear counterstaining (blue). Arrowheads indicate cells expressing the NS3-5B/GFP polyprotein. Bar, 20 μm.
(B) Quantitation of PI(4)P fluorescence from HCV-expressing cells (open symbols) versus HCV-nonexpressing cells (closed symbols). Each point denotes the integrated fluorescence signal from a single cell with the mean fluorescence indicated by the black lines.
(C) Immunostaining of JFH-1 infected Huh7.5.1 cells stably expressing the indicated shRNAs for PI(4)P (red) and GFP (green) with DAPI nuclear counterstaining (blue). Pr, Pearson’s correlation coefficients; bar, 10 μm.