Nrf2 ablation and L2HG accumulation affect the chromatin structure of target genes. (A) ChIP-seq analysis of the enrichment of H3K27Me3 around gene promoters (± 3kb) in bone marrow Ter119⁺ erythroid cells of SS/Nrf2^+/+ and SS/Nrf2^–/– mice. (B) ChIP-seq data tracks for H3K27Me3 enrichment at representative antioxidant (Nqo1), iron/heme metabolism (Hmox1 and Ftl1) and L2hgdh gene loci in the bone marrow Ter119⁺ erythroid cells of SS/Nrf2^+/+ and SS/Nrf2^–/– mice. A schematic representation of genes is shown below the panels, with the blue arrow indicating the TSS and gene orientation. The data are representative of biological triplicate samples. (C-G) ChIP-PCR analysis of the association of Nrf2 (C), H3K27Me3 (D), Tbp (E), and Pol II (F) with the promoter regions of antioxidant (Nqo1, Cat, Gclc, and Gstt1) and iron/heme metabolism (Hmox1, Fth1, Ftl1, Slc7a11, and Slc40a1) related genes in SCD mouse bone marrow Ter119⁺ cells after normalization to the input. Normal rabbit immunoglobulin G was used as an antibody control (G). (H-J) ChIP-PCR analysis of the association of H3K27Me3 (H), TBP (I), and RNA Pol II (J) with antioxidant (NQO1) and iron/heme metabolism (HMOX1, FTH1, FTL1, and SLC7A11) gene loci in erythroid progenitor from patients with SCD cells after TFMB-L2HG (250 μM) treatment. (K) Immunoglobulin G was used as an antibody control. Data represent mean ± SD of 3 biological replicates (n = 3). ∗P < .05. For panels C-F and H-J, a one-way ANOVA with Bonferroni multiple comparison test was used for statistical analyses.