SUV4-20h1 induced epigenetic modification of the γ-globin genes is developmentally specific. (A) Quantitative reverse-transcriptase (Q-RT)–PCR analysis of γ-, β-, and α-globin gene expression in cord blood (CB) and bone marrow (BM). (B) H4K20me3 at the γ-promoter was measured by ChIP in chromatin fractions from erythroid progenitors from CB and BM. (C) Localization of H4K20me3 across the β-globin locus measured by ChIP in chromatin fractions from erythroid progenitors from BM. The precipitated DNA was amplified with primers specific for the indicated regions of the β-globin locus.⁸  HS, hypersensitive site; pro, promoter; ^G/Aγ, intergenic region between ^Gγ- and ^Aγ-globin genes; β-ex3, exon 3 of the β-gene. The error bars correspond to the SD. Each experiment was performed 3 times independently. Normal rabbit IgG served as the control. (D) Western blot analysis of proteins immunoprecipitated from erythroid progenitors from BM with α-PRMT5 Ab (left panel) or α-DNMT3A Ab (right panel). A mock IP with normal rabbit IgG was used as a negative control. Abs used are indicated on the right. (E) Expression of SUV4-20h1 and (F) PRMT5 was analyzed in erythroid progenitors from BM expressing specific shRNAs or scrambled control sequences (scr). (G) Analysis of γ-, β-, and α-globin gene expression in SUV4-20h1-kd, PRMT5-kd and scr control BM cells. The error bars correspond to the SD. Each experiment was performed 4 times independently. (H) Comparison of γ-globin gene expression levels in CB and SUV4-20h1-kd, PRMT5-kd and scr control BM cells by Q-RT-PCR.