Fig. 4.
Induction of autophagy by BECN1 plasmid impairs m6A modification inhibition-induced resistance to HSC ferroptosis. (A) BECN plasmid and METTL4 shRNA or FTO plasmid were transfected into HSC-T6 and HSC-LX2 cells and trested with erastin (10 μM) for 24 h. Real-time PCR were used to measure the mRNA levels of BECN1, MAPILC3B and SQSTM1 (*, p < 0.05, **, p < 0.01, compared with Control group, ##, p < 0.01, compared with BECN1 plasmid group, n = 3 in every group). (B) Western blot showed the protein expression of p62 and LC3-I/II (n = 3 in every group). (C) The endogenous LC3 levels were measured by immunofluorescence (n = 3 in every group). (D) HSC-LX2 and HSC-T6 cells transfected with BECN plasmid and METTL4 shRNA or FTO plasmid were treated with erastin (10 μM) or sorafenib (10 μM) for 24 h. Cell viability was assayed by Cell Counting Kit-8 (n = 3 in every group, *, p < 0.05). (E, F) MDA production, Iron accumulation, GSH depletion and lipid ROS level were assayed by commercial kits (*, p < 0.05, n = 3 in every group).