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. 2018 Mar 30;7(5):2021–2033. doi: 10.1002/cam4.1468

Figure 5.

Figure 5

Doxorubicin promotes the replication of HBV harboring either of two conserved EP elements. (A) The reference sequences of the twelve HBV genomes of genotypes A–D were analyzed by a phylogenetic tree analysis. The accession numbers of the twelve HBV genomes were as follows: genotype A: EU859939, GQ477461, and AB116077; genotype B: GQ205440, JQ688405, and DQ448619; genotype C: AB697490, GQ205441, and AB014367; genotype D: JN257185, U95551, and AB104709. (B) The sequences of the EP element located in enhancer I of the different HBV genotypes were analyzed by MegAlign software (DNASTAR, Madison, WI). (C) The palindromic sequences of the two conserved EP elements among HBV genome genotypes A–D. HepG2 cells were transfected with 2 μg pGEM‐1.3 × HBV expression plasmid (genotype A) or pBB4.5‐1.2 × HBV expression plasmid (genotype C) using lipofectamine 2000 in a 35‐mm dish. After 24 h of transfection, the cells were treated with 1 μmol/L Dox for 1 h. The relative levels of 3.5 kb HBV RNA (D) and total HBV RNA (E) in cells were quantified by qRT‐PCR at 48 h post‐Dox treatment. ACTB mRNA was used as an internal control. According to the above treatment, the relative levels of 3.5 kb HBV RNA (F) and total HBV RNA (G) in HuH7 cells were quantified by qRT‐PCR.