Fig. 3

ZNF148 inhibits the transcriptional activity of HBV cccDNA. (A-B) HepG2-NTCP cells were infected with HBV and were then transfected with the vector/ZNF148 plasmid or infected with lentivirus expressing shCont/shZNF148. After 5 d, the cells were treated with actinomycin D (5 μg/ml) and harvested at the indicated times. (C-D) The cells were treated with 0.2 mM EU and harvested after 24 h. The amount of newly synthesized EU-labeled RNA was quantified via real-time PCR. (E-F) The ratios of total RNA and 3.5-kb RNA to cccDNA are considered markers of HBV cccDNA transcriptional activity. The level of HBV cccDNA was measured by real-time PCR. (G-H) Luciferase reporter plasmids containing the HBV promoter sequence were cotransfected with the vector/ZNF148 plasmid or transfected in combination with infection with lentivirus expressing shCont or shZNF148 into Huh7 cells. After 36 h, luciferase activity was measured via a dual luciferase assay. *P < 0.05, **P < 0.01