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. 2018 Dec 5;42(1):67–78. doi: 10.14348/molcells.2018.0255

Fig. 2. HBx stimulates HBV replication via DNA methylation.

Fig. 2

(A) HepG2 cells were transfected with 1.2-mer WT as described in Fig. 1 in the presence of an increasing concentration of 5-Aza-2′dC. For the determination of viable cell number, MTT assay was performed as described in Methods (n = 4). Levels of extracellular HBV particles were determined by IP-coupled conventional PCR. (B) HepG2 cells were transfected with either 1.2-mer WT or its HBx-null derivative for 48 h in the presence or absence of 5 μM 5-Aza-2′dC. Levels of extracellular HBV particles were determined by HBsAg ELISA (n = 5). (C) Levels of intracellular HBV DNA and (D) extracellular virus particles from HepG2 cells prepared as in (B) were determined by IP-coupled real-time PCR (n = 6). (E) HepG2 cells were transfected with either 1.2-mer WT or its HBx-null derivative along with the indicated amount of DNMT1 shRNA plasmid or SC shRNA plasmid for 48 h, followed by Western blotting. (F) Levels of extracellular HBV particles released from HepG2 cells prepared in (E) were determined by IP-coupled real-time PCR (n = 6).