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. 2012 Sep 17;109(45):18471–18476. doi: 10.1073/pnas.1204668109

Fig. 3.

Fig. 3.

Interaction between HBx and Bcl-2 proteins is critical for HBV DNA replication in human hepatic cells. (A) Southern blot analysis of HBV DNA replication in HepG2 cells transfected with the wild-type or mutant (G124L, I127A) pHBV replicon 3 d posttransfection. rc/ds DNA represents relaxed circular DNA and double-stranded DNA. ssDNA represents single-stranded DNA. (B) Measurement of the levels of the HBV core protein (HBcAg) in transfected HepG2 cells. The difference between cells transfected with the wild-type and mutant (G124L, I127A) pHBV is shown as fold change to wild type. The data represent mean ± SD from three independent experiments. ***P < 0.0001. (C) Northern blot analysis of HBV transcription in HepG2 cells transfected with the wild-type or mutant pHBV replicon 3 d posttransfection. Different HBV mRNAs are indicated. GFP mRNA from a cotransfection marker and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA were used as controls. (D) Quantitative PCR analysis of HBV DNA replication in HepG2 cells transfected with the wild-type or mutant pHBV replicon with or without 5 μM ionomycin treatment. Three days posttransfection, cytoplasmic HBV viral particles were isolated and the viral DNA replication intermediates were quantified by real-time PCR (Materials and Methods). The results represent the fold change of the replicative intermediates from the mutant pHBV replicon compared with those from the wild-type pHBV replicon in HepG2 cells, using two different primer sets (one specific to the HBS ORF and one specific to the polymerase ORF). Data are presented as mean ± SEM. At least three independent experiments were performed for each dataset. ***P < 0.0001.