Fig. 5

ZNF148 regulates the HBV life cycle by suppressing the binding of RXRα to HBV ENII/Cp. (A-B) The impact of ZNF148 on the recruitment of RXRα to HBV ENII/Cp was evaluated via a ChIP assay. The promoter of GAPDH served as the control in this analysis. The results are presented as percentages of input. (C) The binding sites and mutations are shown in the HBV 1.3 genome. (D) The vector/ZNF148 plasmid was cotransfected with the indicated luciferase reporter plasmids into HepG2-NTCP cells. After 36 h, the luciferase activities of pGL3-ENII/Cp and pGL3-ENII/Cp Mut (with mutation of the RXRα binding sites) were measured by a dual luciferase assay. (E-G) HepG2-NTCP cells were transfected with the HBV1.3 genome or the HBV1.3 genomic mut construct (with mutation of the RXRα binding sites); after 24 h, the vector/ZNF148 plasmid was transfected into the cells. The levels of total HBV RNA, 3.5-kb HBV RNA, and HBV core DNA were measured via RT-PCR. (H) The ZNF148 or RXRα plasmid was cotransfected with the indicated luciferase reporter plasmids into HepG2-NTCP cells, and luciferase activity was measured using a dual luciferase assay. (I-K) HepG2-NTCP cells were transfected with the ZNF148 or RXRα plasmid, and HBV RNA levels were measured using real-time PCR and Northern blotting. (L-M) The HBV core DNA level was measured using real-time PCR and Southern blotting. (N) The HBc protein level was measured by Western blotting. *P < 0.05, **P < 0.01