FIG 1.

Construction of E1-mCherry-labeled HCV Jc1 variants. (A) Schematic representation of HCV-Jc1 genomes generated in this study. The HCV-Jc1 genome is depicted at the top for reference. We modified this backbone to express mCherry within the E1 glycoprotein with or without NS5A-GFP, Flag-E2, or the A4 epitope in E1 for recognition by the anti-E1 A4 antibody (see Materials and Methods for details). Green box, GFP; red box, mCherry; blue box, Flag tag. (B) Huh7.5 cells were lysed at 48 h postelectroporation (hpe) with RNA of Jc1-WT or Jc1-ΔE1/E2 or variants of Jc1-E1(A4), and expression of viral proteins was analyzed by Western blotting. Blots representative of results of at least three individual experiments are shown. (C) Confocal images of HCV-Jc1-E1(A4) and HCV-Jc1-E1(A4)-mCherry Huh7.5 cells electroporated at 48 hpe show subcellular localization of viral proteins visualized by immunofluorescence staining and localization of E1-mCherry. Scale bar, 35 μm.