Fig 7.
HCV–co-opted host proteins VAP-A and VAP-B are enriched in DMVs. (A) Purified ER control (a and b) and HCV-remodeled membranes (c and d) were consecutively labeled for NS5A (15-nm gold) and for VAP-A (a and c) or VAP-B (b and d) (10-nm gold). After negative staining, they were examined by TEM. White and black arrowheads highlight 10- and 15-nm gold particles, respectively. Scale bars represent 100 nm. (B) Quantification of gold particles on membranes. At least 100 gold particles were counted, and the number of gold particles associated with membranes per area of the grid is depicted. (C) Number of single- and double-labeled structures containing either NS5A or VAP-A/B, or both viral and cellular protein, respectively, per area on the grid. At least 50 membranous structures per condition were analyzed. (D) Amount of 10-nm colloidal gold particles per micrometer of membrane length. Only membranes positive for NS5A were considered for the analysis of NS4BHA-captured membranes. The analysis is based on ∼50 membranous structures with a total membrane length of more than 50 μm for each condition.