GATA factors and FOG1 regulate MC genes in erythroid cells. (A) mRNA levels of indicated genes as determined by quantitative RT-PCR of G1E cells, and G1E-ER4 and G1E(V205M)-ER cells treated with estradiol for 20.5 hours. Data were normalized to actin and plotted as fold change compared with uninduced G1E cells; n = 3-5. *P < .001; **P < .05; ns indicates not significant. (B) Overexpression of GATA2 in G1E-ER4 cells (G1E-ER4 + GATA2) stimulates MC gene expression, whereas estradiol-activated GATA1-ER (+est.) represses them; n = 6. *P < .05 comparing G1E-ER4 + GATA2 ± estradiol treatment. (C-G) ChIP for GATA2 (C-D), GATA1 (E-F), and FOG1 (G) at FcerIb and Cpa3 in G1E cells (C,E,G) or BMMC (D,F). Control regions included: −7 kb upstream of the FcerIb promoter (UR), an intronic region 3 kb downstream of the Cpa3 promoter (5′TR), the 5′transcribed region of Cd4 (Cd4 5′TR), and a region −224.9 kb upstream of the Kit gene (UR). Positive control, the GATA1-activated Eraf erythroid gene. n = 3-8 independent ChIP experiments per primer set. (H-I) ChIP against MTA2 (H) and RbAp46 (I) in G1E cells and G1E-ER4 (+est.); n = 4-6. The promoter of GATA1-activated Hbb-b1 served as a positive control. *P < .01, #P < .05 by Student t test compared with IgG controls (C-I). (J) ChIP for acetylated histone H3 (acH3) in G1E cells, and G1E-ER4 and G1E(V205M)-ER cells treated with estradiol for 20.5 hours; n = 4-6. *P < .01, #P < .05 by Student t test. Error bars represent SEM.