FIG 4.

WTAP negatively regulates HCV virion production. (A and B) Focus-forming assay of supernatant harvested from HCV-infected (48 h, MOI of 0.3) Huh7 cells depleted of indicated proteins (A), as well as immunoblot analysis of these lysates (B). (C and D) Focus-forming assay of supernatant harvested from HCV-infected (48 h, MOI of 0.3) Huh7 cells overexpressing the indicated proteins (C), as well as immunoblot analysis of these lysates (D). For WTAP and METTL3, protein-specific antibodies detect both endogenous and overexpressed proteins. (E) Gaussia luciferase values from the supernatant of Huh7.5-CD81 KO cells treated with the indicated siRNA and transfected with a full-length HCV RNA containing a Gaussia luciferase reporter cassette, measured at indicated hour posttransfection. Graphs show mean ± SD; n = 3 (C and E) or 4 (A) biological replicates. Blots are representative of 3 independent experiments. Data were analyzed by one-way ANOVA with Šidák’s multiple-comparison test (*, P < 0.05; ***, P < 0.001).