Fig. 6.

Nup98 interacts with Core and relocalizes to cLDs in infected cells. (A) Cell lysates from naive and clone 2-infected Huh-7.5 cells (6 dpi) were immunoprecipitated (IP) with anti-Nup98 Ab. Beads without Ab (control beads) or conjugated to isotype-matched IgG served as negative controls. Immunoblot analysis with anti-Nup98 and anti-Core Abs is shown. (B) Huh-7.5 cells infected with clone 2 for 4 d were cryosectioned and stained with Abs to Core (a), Nup98 (b and d), or both (c). Representative iEM images are shown. (a) Core is enriched on cLDs juxtaposed to endoplasmic reticulum (ER) membranes. Membrane curvature may represent a nucleocapsid budding event (white arrow). Spherical particles within vacuoles may be HCV virions at early maturation stage (stars). (c) Costaining with Core-specific (12 nm) and Nup98-specific (6 nm) Abs. (d) Low-magnification image shows the expected localization of Nup98 on both sides of the nuclear envelope, in the central channel of the NPC (Inset), and the additional presence of Nup98 on cLD. (Scale bars, 100 nm.)