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. 2021 Dec 17;298(2):101501. doi: 10.1016/j.jbc.2021.101501

Figure 2.

Figure 2

Octyl-L-2HG increases HIF-1α activity in macrophages. LPS (24 h) induced HIF-1α and pro-IL-1-β in BMDMs pretreated for 3 h with octyl-L-2HG (A), or octyl-D-2HG (B). HIF-1α as detected in cells treated with octyl-2HG as indicated (C) or with 500 μM octyl-2HG (D). LPS (24 h) induced HIF-1α and pro-IL-1β in BMDMs pretreated for 3 h with octyl-2HG at the indicated concentration (E) or treated simultaneously with octyl-2HG (500 μM) and LPS (F). Expression of Il1b, Phd3, Nos2, and Glut1 in LPS stimulated (24 h) BMDMs pretreated with (G) octyl-L-2HG or (H) octyl-D-2HG (500 μM, 3 h), n = 8 from three independent experiments. TNF-α (I) in BMDMs treated with octyl-D-2HG or octyl-L-2HG (500 μM for 3 h), stimulated with LPS (3 h), n = 3. Glycolysis and glycolytic capacity (fold over unstimulated control) in octyl-D-2HG or octyl-L-2HG (500 μM) treated BMDMs (J), n = 3. ECAR in unstimulated and octyl-L-2HG (500 μM) (K), or octyl-D-2HG (L) treated BMDMs. Western blots are representative of n = 4 (A–B), or n = 3 (C–F), graphs represent the mean ± S.E.M.; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.001; ns, not significant; one-way ANOVA. 2HG, 2-hydroxyglutarate; BMDM, bone-marrow-derived macrophage; LPS, lipopolysaccharide.