Figure 4.

Decitabine treatment reduces NFE2 expression by reversing H3K9 hypomethylation and H3Y41 phosphorylation. (A) Effect of decitabine treatment on the expression of NFE2, of JMJD1C, and on total H3K9me2 levels in HEL cells. HEL cells were treated with 400 ng/μL decitabine or dimethyl sulfoxide (DMSO). Lysates were interrogated by western blot. (B-D) Effect of decitabine treatment on H3K9me2 levels, HP1α binding, and H3Y41 phosphorylation at the NFE2 locus in HEL cells. Top panels: HEL cells were treated as in panel A and chromatin immunoprecipitated with an antibody against H3K9me2 (B), HP1α (C), H3Y41ph (D), or an IgG control as indicated. Chromatin immunoprecipitated DNA was amplified with the primers illustrated in Figure 3C. One representative result of at least 3 independent experiments is shown. Bottom panels: densitometric analysis of the ChIP results. Graphs represent mean ± SEM. *P < .05, **P < .01 by the Student t test. DAC, decitabine.