Fig 4.

Effects of mutations in the core-E1 signal peptide from GT3a on virus production and proteolysis of core. (A) HuH-7 cells were electroporated with in vitro-transcribed RNA, and cell supernatants were harvested at 24-h intervals. Virus titers were determined by measurements of TCID₅₀ values. Data shown represent the average values from three independent experiments and are for the following constructs: JFH1-GT3a-wt1 (□), JFH1 (■), JFH1-GT3a-mut1 (○), and JFH1-GT3a-mut2 (●). Error bars indicate standard deviations. (B and C) HuH-7 cells were electroporated with in vitro-transcribed RNAs as indicated. (B) Cell extracts were harvested at 72 h and analyzed by Western blotting with antibodies against core and NS5A. (C) Cells were prepared for immunofluorescence at 72 h after electroporation and analyzed by confocal microscopy with antibodies against core and ADRP. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), and the scale bar represents 10 μm.