Figure 6
Figure 6. NRF2 and KLF2 bind the β-globin LCR in differentiating primary human erythroid cells. (A) NRF2 ChIP analysis of day 13 CD34+ cells during erythroid differentiation. Cells were treated with 1 and 5μM tBHQ on days 9, 11, and 13. (B) KLF2 ChIP of CD34+ cells treated with 5μM statin on day 5. Cells were harvested for cross-linking and ChIP analysis on days 8, 12, 15, and 16 of differentiation. qPCR was performed on immunoprecipitated DNA using primers that amplify HS3, HS2, γ-globin, and the β-globin promoter. The necdin promoter was used as a negative control and the NQO1 promoter was used as a positive control for the NRF2 ChIP. Results are expressed as the fold enrichment over IgG. (C) Comparison of KLF2 mRNA levels and KLF2 protein bound to HS3 and HS2. The data are presented on the same graph to illustrate KLF2 binding at specific points during differentiation compared with KLF2 gene expression. Error bars represent ± SEM. *P < .05 compared with 0μM concentration of drug. NS indicates not significant by t test (P < .05, n = 2).

NRF2 and KLF2 bind the β-globin LCR in differentiating primary human erythroid cells. (A) NRF2 ChIP analysis of day 13 CD34+ cells during erythroid differentiation. Cells were treated with 1 and 5μM tBHQ on days 9, 11, and 13. (B) KLF2 ChIP of CD34+ cells treated with 5μM statin on day 5. Cells were harvested for cross-linking and ChIP analysis on days 8, 12, 15, and 16 of differentiation. qPCR was performed on immunoprecipitated DNA using primers that amplify HS3, HS2, γ-globin, and the β-globin promoter. The necdin promoter was used as a negative control and the NQO1 promoter was used as a positive control for the NRF2 ChIP. Results are expressed as the fold enrichment over IgG. (C) Comparison of KLF2 mRNA levels and KLF2 protein bound to HS3 and HS2. The data are presented on the same graph to illustrate KLF2 binding at specific points during differentiation compared with KLF2 gene expression. Error bars represent ± SEM. *P < .05 compared with 0μM concentration of drug. NS indicates not significant by t test (P < .05, n = 2).

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