Figure 4.

G9a mediates H3K9me2 at ERO1α promoter in hepatocytes with Hcy treatment. (a and b) The H3K9me2 levels in liver tissues of cbs ^{+/ −} mice (n = 6) and hepatocytes treated with Hcy (n = 3) were analyzed by western blot. (c) The enrichment of H3K9me2 on ERO1α promoter was assayed by ChIP assay in hepatocytes after Hcy treatment (n = 6). (d and e) G9a protein expression was detected by western blot in liver tissue of cbs ^{+/ −} mice (n = 6) and hepatocytes treated with Hcy (n = 3). (f) G9a protein expression was measured by western blot in hepatocytes transfected with G9a‐overexpressed plasmid (pG9a) (n = 3). (g) Protein expression of G9a was examined by western blot in hepatocytes after transfection with three G9a siRNAs (si‐G9a‐1/2/3) (n = 3). (h) ChIP assay for analysis of the enrichment of H3K9me2 on ERO1α promoter in hepatocytes after transfection with pG9a or si‐G9a in the presence of Hcy (n = 3). (i) Western blot was used to measure the protein expression of ERO1α in Hcy‐treated hepatocytes after transfection with pG9a and si‐G9a (n = 3). Data represents mean ± SD, *p < 0 .05, **p < 0 .01. ChIP, chromatin immunoprecipitation; ERO1α, endoplasmic reticulum oxidoreductase 1α; Hcy, homocysteine; siRNA, small interfering RNA