Figure 2
HBx increases methylation of the human MTA1 promoter CpG island1. (a) Representative sequencing traces from bisulfite-modified direct sequencing. ▾ denotes methylated or unmethylated cytosines. Seven of unmethylated cytosines were converted to uracil in empty vector (EV)-transfected WRL-68 cells, but two in p3XFLAG7.1-HBx-transfected cells. (b) p3XFLAG7.1-HBx or EV was transfected into the indicated cells or treated with vehicle or 2 mg/ml of doxycycline (Doxy) for 24 h. Results from bisulfite sequencing analysis of the MTA1 promoter CpG island1 were shown. Filled squares, methylated; and open squares, unmethylated. Numbers at the bottom indicate the proportion of methylated CpG sites relative to total CpG sites examined. (c) Indicated amount of p3XFLAG7.1-HBx or EV was transfected into cells, or treated with doxycycline. Whole-cell lysates were prepared and protein levels were evaluated by western blotting.
