Figure 4.

ZNF143 modulates HBV replication through regulating HBV core promoter activity in hepatic cell lines. (A–F) Huh7 cells were co-transfected with plasmid pSM2 and specific siRNAs against ZNF143 (siZNF143) at 40 nM (A, C, E) or plasmid Flag-ZNF143 (B, D, F), and harvested at 72 h post-transfection. Secreted HBsAg (A, B) and HBeAg (C, D) levels in culture supernatants were determined by chemiluminescence immunoassay. (E, F) Encapsidated HBV replicative intermediates were isolated and detected by Southern blotting. (G–J) Huh7 cells were co-transfected with siZNF143 or negative control (siNC) at 40 nM and HBV promoter luciferase reporters containing the region of pCP for 48 h with Renilla as an internal control. The data from dual-Glo luciferase report assay were calculated by fold change and normalized to the siNC samples. HepG2.2.15 (I) and Huh7 cells (J) were separately treated as (A–F). At 72 h post-transfection, western blotting analysis was performed to detect the levels of ZNF143, FXRα, and HBV core proteins using beta-actin as a loading control. *P < .05; **P < .01; ns, no significance. RC, relaxed circular DNA; SS, single-stranded DNA.