Cited2 deficiency affects the expression of HSC quiescence related genes. (A) Gene-expression analysis by real-time PCR. CD34⁻ LSK cells were sorted for RNA isolation from each genotype. The analysis was repeated by 3 or 4 independent experiments. Two mice per genotype were used in each experiment. Data are shown as relative expression compared with WT controls (means ± SEM). (B-C) HIF-1α and HDAC1 bind to the Hes1 promoter. (B) Relative expression of Hes1 in EML C1 cells treated with DFO (100μM) with or without suberoylanilide hydroxamic acid (2μM). (C) ChIP assay in EML C1 cells was performed with anti–HIF-1α, anti-HDAC1, and IgG Abs. The precipitated DNA was subjected to quantitative real-time PCR with primers amplifying nt −235 to −1 of the Hes1 promoter. (D-E) Cited2 and Smad2/3 are recruited to the Egr1 promoter. (D) Relative expression of Egr1 in starved EML C1 cells after treatment with TGF-β1 (7.5 ng/mL) for the indicated time periods. (E) Starved EML C1 cells were treated with TGF-β1 for 1 hour and used for ChIP assay with anti-Smad2/3, anti-Cited2, and IgG Abs. The precipitated DNA was subjected to quantitative real-time PCR with primers amplifying nt −2342 to −1965 of the Egr1 promoter. These experiments were repeated 3 times. *P < .05; **P < .01. NS indicates not significant.