CARM1 represses γ-globin expression.
A, Western blot analysis of indicated protein levels in cell lysates from the scramble control (Scr), CARM1-KD1, and CARM1-KD2 Lys-562 cells. GAPDH and histone H4 were used as loading controls. Blots are representative of three independent experiments. B and C, quantitative real-time PCR analysis of CARM1 (B) and γ-globin (C) mRNA level in Scr, CARM1-KD1, and CARM1-KD2 Lys-562 cells normalized to β-actin mRNA. The results are shown as the mean ± S.D. from three independent experiments. Two-tailed Student's t-tests were used to compare means. **, p < 0.01 compared with the Scr control. D, ChIP analysis of H3R17me2a and H4R3me2s enrichment at the γ-promoter in Scr, CARM1-KD1, and CARM1-KD2 Lys-562 cells. IgG was used as the negative control. The results are shown as the mean ± S.D. from three independent experiments. Two-tailed Student's t-tests were used to compare means. **, p < 0.01 compared with the Scr control.