Fig. 5.
dL2HGDH activity is inhibited by lactate. (A–C) 2HG levels in mid-L2 larvae raised on semidefined media containing 0, 5, or 10 mM lactate. Dietary lactate induced elevated 2HG levels in both w1118 controls and dLdh16/dLdh17 (dLdh−) mutants, but not dL2HGDH12/dL2HGDH14 (dL2HGDH−) mutants. (D) Ability of lactate to inhibit L-2HG degradation was assessed by incubating partially purified dL2HGDH with 2 mM L-2HG and 0, 2, or 10 mM lactate (n = 6, data presented as mean ± SD). (E) Relative 2HG levels in mid-L2s of w1118 controls, dL2HGDH12/14; dLdh16/17 single mutants, and dL2HGDH12/14; dLdh16/17 double mutants. (F) Relative abundance of D-2HG and L-2HG levels in dL2HGDH; dLdh mutant midsecond-instar larvae was measured using GC-MS. In A–C and E, data are represented as box plots (n = 6). *P < 0.05; **P < 0.01; ***P < 0.001.