DNA methylation inhibits integrated HBV DNA transcription. (A) genome-wide methylation levels of integrated HBV DNA and host genome.CG, CHG and CHH represent the three forms of C nucleotide distribution on the genome (H represents A, T or C nucleotide). (B) Heat map of host genome methylation levels around integration sites in the PLC/PRF/5 cell line. (C) Correlation between host genome methylation levels within 500 bp upstream and downstream of HBV integration sites and the transcriptional level of integrated HBV DNA. (Methylation levels of genome around Integration sites on chromosome 4 and chromosome 8 with severely outlying were excluded from the correlation analysis.) The y-axis represents log2-transformed HBV chimeric read counts from each integration site, indicating the transcriptional level of each integration site. (D) RT-qPCR was used to detect the relative expression of intracellular HBV RNA of PLC/PRF/5 cells before and after AzaD treatment using primer that targeting S region (329-451 nt) of HBV. The control group and experimental group were treated with DMSO and different concentrations of AzaD, respectively, for 48 h before harvesting. (E) Western Blot was used to detect the intracellular HBsAg level of PLC/PRF/5 cells before and after AzaD treatment. (F) Quantitative PCR detection of transcription levels at each HBV integration sites in PLC/PRF/5 cells before and after AzaD treatment using primer that targeting the specific sequence of each integrated sites (Table S1). The expression level of genes was normalized using β-actin. * Indicates P < 0.05, **** indicates P < 0.0001.