HBx regulates miR-125a transcription in a methylation-dependent manner.

<p>(A) L-O2, L-O2-pCMV, and L-O2-HBx cells were analyzed for miR-125a expression by real-time RT-PCR. (B) L-O2-HBx cells were transfected with control siRNA or HBx siRNA, and miR-125a expression was analyzed by real-time RT-PCR. (C) The methylation of miR-125a CpG sites was analyzed in L-O2 and L-O2-HBx cells by bisulfite sequencing analysis. At least five independent clones were sequenced per sample. Open and filled circles represent nonmethylated and methylated CpG sites, respectively. (D) L-O2 cells were analyzed for miR-125a expression by qRT-PCR after treatment with 5 μM 5′-Aza for 72 h. Data are from three independent experiments and are presented as the mean ± SEM (**P<0.01, ***P<0.001).