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. Author manuscript; available in PMC: 2016 Aug 4.
Published in final edited form as: Cell Metab. 2015 Jul 23;22(2):291–303. doi: 10.1016/j.cmet.2015.06.021

Figure 3. Potential Mechanisms of L2HG Accumulation in Hypoxia.

Figure 3

(A) Enzymatic and cofactor requirements for L2HG metabolism. (B and C) Intracellular 2OG (B) and 2HG (C) in normoxia and hypoxia measured after treatment with a cell-permeable 2OG analogue, TFMB-2OG. (D and E) 2OG (D) and 2HG (E) in cells treated with the OGDHC inhibitor KMV (20 mM) ± 0.2% oxygen. (F) Correlation between 2OG and 2HG from experiments illustrated in B-E. (G) Immunoblot of MDH1 and MDH2 in siRNA-treated LF. (H–J) 2OG (H), 2HG (I), and 2OG/2HG (J) in MDH knockdown cells. (K) Relative changes in mRNA levels of 2HG metabolic enzymes in response to hypoxia. (L) Immunoblot of LF cell lysates of proteins involved in L2HG metabolism. (M) Intracellular malate determined by targeted LC-MS. (M) Cellular NADH/NAD+ determined by enzymatic cycling assay. Data are mean ± SEM. See also Figure S3.