Nrf2 regulates L2HG production to affect histone methylation. (A) S-adenosyl methionine levels in spleen Ter119⁺ cells from SS/Nrf2^+/+ and SS/Nrf2^–/– mice. (B) Quantitative PCR determined the expression of selected core histone methylases and demethylases in spleen Ter119⁺ cells from SS/Nrf2^+/+ and SS/Nrf2^–/– mice. (C) GC-MS analysis of metabolite levels in SS/Nrf2^+/+ and SS/Nrf2^–/– mouse spleens (top 25 metabolites with differences are presented). (D) Relative levels of 2HG and 2OG in SS/Nrf2^+/+ and SS/Nrf2^–/– mouse spleens. (E-F) Levels of L2HG and D2HG (E) and their ratios with respect to 2OG (F) in SS/Nrf2^+/+ and SS/Nrf2^–/– mouse spleens after normalization to protein content. (G) Effect of cell-permeable 2OG (DMOG), D2HG (octyl-D-2HG) and L2HG (TFMB-L2HG) on H3K9Me3 and H3K27Me3 levels in EPs from patients with SCD. EPs were cultured for 12 days with H₂O₂ (200 μM)/hypoxia (1% O₂) and treated with the indicated concentrations of DMOG, octyl-D-2HG, or TFMB-L2HG, and histone methylation levels were determined in the histone extracts. (H) Total 2HG, D2HG, and L2HG levels in PBMNs from HD and patients with SCD. Data represent mean ± SD (n = 6) (A-F,H) or 3 biological replicates (G). ∗P < .05. DMOG, dimethyl 2OG; TFMB-L2HG, trifluoromethylbenzyl ester derivative of L2HG.