H3K9Me2 levels decrease during erythroid differentiation and are further decreased by EHMT1/2 pharmacologic inhibition or knockdown in primary adult human erythroid cells. (A) EHMT1 and EHMT2 mRNA expression in human CD34⁺ cells from adult or umbilical CB. Results show the mean value ± SD of 2 different donors for adult CD34⁺ and 2 pools of 10 cords for CB CD34⁺. (B) Western blot analysis showing EHMT1, EHMT2, and lamin B1 (loading control) in primary adult human cells during erythroid differentiation ex vivo. (C) Representative flow plots of H3K9Me2 level according to the cell number and the size during erythroid differentiation ex vivo. (D) Representative histogram showing the fluorescence intensity for H3K9Me2 relative to the cell number in erythroid cells differentiated in the presence of 0.25 μM UNC0638 or the vehicle control for 7 days. (E) Dot plots showing the median fluorescence intensity (MFI) for H3K9Me2 after 7 days of treatment according to the UNC0638 concentration (mean ± SD, n = 8 biological replicates). (F) Dot plots showing the MFI for H3K9Me2 in EHMT1 or EHMT2 knockdown cells (mean ± SD, n = 5-8 biological replicates). ***P < .001.