Figure 5. OSBP localization to membranous web requires PI4KA activity.
(A) The NS3-5B(NS5A/GFP) polyprotein was expressed in Huh7.5.1/T7 cells stably expressing a nontargeting or a PI4KA shRNA. 48 hr post-transfection, cells were immunostained for OSBP (red) and NS5A (green) with DAPI nuclear counterstaining (blue). M1 and M2, Manders’ coefficients; M1 represents the fraction of NS5A overlapping OSBP. Bar, 10 μm.
(B) The NS3-NS5B(NS5A/GFP) polyprotein was expressed in Huh7.5.1/T7 cells. 48 hr later, cells were treated with 0.1% DMSO or 4 μM AL-9 for 24 hr, and then immunostained for NS5A (green) and endogenous OSBP (red) with DAPI nuclear counterstaining (blue). M1 and M2, Manders’ coefficients; M1 represents the fraction of NS5A overlapping OSBP. Bar, 10 μm.