The impact of E2-p7 processing on NS2 localization and its interaction with NS3 from the following in panel A: frame a, HJ3-5; b, HJ3-5/p7(KRAA); c, HJ3-5/E2(AR); d, HJ3-5/E2/IRES; e, HJ3-5/E2/IRES/p7(KRAA); f, HJ3-5/HAp7; and g, HJ3-5/HAp7(KRAA). Confocal image analysis using an Olympus FluoView FV1000 laser scanning confocal microscope of cells at day 2 postelectroporation with HCV RNAs encoding indicated genomes. Anti-NS2 antibody (green) and LipidTOX deep red neutral lipid stain (red) were used to detect NS2 and lipid droplets. The numbered images at the bottom are enlargements of the corresponding areas from the images at the top. (B) The relative percentages of NS2-positive cells displaying different NS2 localization patterns NS2 localization patterns were analyzed from 50 NS2 immunostaining-positive cells from two independent (lanes a, b, c, d, and e) or single (lanes f and g) experiments. Punctate, NS2 localization similar to that detected from frames a, d, f, and g in panel A; intermediate, NS2 localization similar to that observed from frame e' in panel A; nonpunctate, NS2 localization similar to that detected from frames b and c in panel A.