Fig. 3

HBx suppresses SLC7A11 expression through H3K27me3 by EZH2. Primary hepatocytes were transduced with lenti-vector or lenti-HBx. A The mRNA levels of GPX4, SLC11A2, SLC7A11 and ACSL4 were determined by qRT-PCR. The enrichments of EZH2 and H3K27me3 on promoters of B SLC7A11, C GPX4, D SLC11A2 and E ACSL4 were determined by ChIP assay. Normal IgG served as a negative control. F The protein levels of EZH2 and H3K27me3 were determined by western blot. G The direct interaction between HBx and EZH2 was assessed by co-IP. Normal IgG served as a negative control, and whole cell lysates served as an input control. H and I Transduced cells were pre-treated with CHX for 1 h. Cells were harvested at 0, 3, 6, 9 and 12 h. The protein level of EZH2 was determined by western blot. J and K The protein level of EZH2 and SLC7A11 were determined by western blot. *, P < 0.05, **, P < 0.01, ***, P < 0.001