Figure 6.

Effect of RN-1 on histone modifications and γ-globin promoter DNA methylation. (A) ChIP analysis of levels of pol II, Histone H3K4Me2, H3K4Me3, H3K9Ac, and H3K9Me2 associated with the ε-, γ-, and β-globin promoters and IVS regions in purified bone marrow (BM) erythroid cells from RN-1 treated baboon pre and post treatment. Anemic baboons were treated with 0.5 mg/kg/d RN-1 for 2d and BM aspirates were obtained 48 h following the last RN-1 treatment. Asterisks (*) designate significant differences (P<0.05) between pre- and post-treatment samples. (B) (Left) Levels of cytosine methylation at three non-polymorphic CPG residues in the 5′ γ-globin promoter region in purified BM erythroid cells from RN-1 treated baboons pre and post treatment. Paired pre and post treatment samples from individual baboons are shown. The horizontal bar shows the mean % dmC value of all samples. Anemic baboons were treated with either 0.5 mg/kg/d (filled triangles, filled squares, filled circles, filled diamonds, lined squares) or 0.25 mg/kg/d (open triangles, open circles). (Right) Changes in γ-globin synthesis in peripheral blood reticulocytes of baboons pre and post treatment. Paired pre and post treatment samples from each individual are shown. The horizontal bar shows the mean level of γ-globin synthesis (γ/γ+β) of all samples.