MTCH2 knockdown increases nuclear levels of pyruvate and nuclear levels of PDH complex. (A) Levels of mitochondrial and nuclear pyruvate were measured from control and MTCH2 shRNA-knockdown clones of OCI-AML2 cells as described in the supplemental Methods (****P < .0001). (B) Levels of intracellular lactate were quantified by nuclear magnetic resonance from control and MTCH2 shRNA-knockdown clones of OCI-AML2 cells, as described in the supplemental Methods. (C-D) Nuclear and mitochondrial fractions were isolated from control and MTCH2 shRNA-knockdown clones of (C) OCI-AML2 or (D) TEX cells and analyzed by immunoblotting. Antibodies against PDH (E1α and E2), VDAC, and histone H3 were used. Representative immunoblots are shown. The relative expression of PDH subunits, E1α and E2 in mitochondria and nucleus, was quantified by densitometry and normalized to loading controls SOD1 (for E1 α), VDAC (for E2), and histone H3, respectively, and was expressed relative to control shRNA. Data are represented as mean ± SD from 3 independent experiments. **P < .01; ***P < .001; ****P < .0001 by ANOVA.