Abundance of 2-hydroxyglutarate is increased in LPS-stimulated macrophages.A, relative metabolite abundance in control and LPS-stimulated (100 ng/ml, 24 h) BMDMs. B, metabolites significantly upregulated (red) or downregulated (blue) following 24 h LPS stimulation, n = 5; FC, Fold change. C, 2HG levels in control and LPS stimulated (24 h) BMDMs, (D) intracellular 2HG concentration, and (E) relative abundance of each enantiomer of 2HG in control and LPS-stimulated (100 ng/ml, 24 h) BMDMs, n = 5. Expression of (F) D2hgdh and (G) L2hgdh in BMDMs stimulated with LPS, n = 9 from three independent experiments. H, relative changes in L-2HGDH protein level quantified with TMT coupled LC-MS, n = 4. I, possible routes of synthesis and degradation of L-2HG and D-2HG. Graphs represent median ± min/max (C); mean ± S.D. (D–E), mean ± S.E.M. (F–G); ∗∗ p < 0.01, ∗∗∗ p < 0.001, unpaired t test. 2HG, 2-hydroxyglutarate; BMDM, bone-marrow-derived macrophage; LPS, lipopolysaccharide.