Figure 3.

MTHFD2 Interacts with Complex III to Regulate Glucose Metabolism in mESCs

(A) Summary of enriched GO terms that were potently upregulated and downregulated by Mthfd2. p < 0.05. For details, see Table S2.

(B and C) Heatmap of DEGs about glycolysis (B) and OXPHOS (C) between control mESCs and Mthfd2 KD mESCs.

(D and E) Examination of intracellular glucose levels (D) and lactate levels (E) in Mthfd2 KD mESCs. Data are pooled from three independent experiments (mean ± SD). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 (Student's t test) compared with the control.

(F and G) CoIP results showing the specific interactions between endogenous MTHFD2 and both UQCRC2 and CYC1 in mESCs (F) and in the cytoplasm fraction of mESCs (G).

(H and I) GST pull-down assays for interaction between UQCRC2-His (H), CYC1-His (I), and MTHFD2-GST fusion proteins at 250 mM NaCl containing GST binding buffer.

(J) Western blot analysis of the levels of the UQCRC2 and CYC1 proteins in Mthfd2 KD mESCs. β-Tubulin was used as a loading control.

(K) Western blot analysis of the levels of UQCRC2 protein in Mthfd2 KD mESCs at indicated times post cycloheximide (CHX) treatment. GAPDH was used as a loading control.

See also Figures S3–S5.