Fig. 7
α-KG induces the activation of the central carbon metabolism. a The results of the immunoblot analysis of phospho-mTOR (Ser2448), mTOR and histone H3 in CD8 T cells cultured under the indicated conditions for 48 h. The protein amount of histone H3 was used as a loading control. b DM-α-KG-dependent induction of the phosphorylation (Ser240/244) of ribosomal S6 protein in CD8 T cells stimulated for 24 h. c, d Naive CD8 T cells were stimulated with anti-TCR-β, anti-CD28 mAbs plus IL-2 without glutamine in the presence or absence of DM-α-KG for 36 h and then ECAR (c) and the OCR (d) were determined. e The results of the quantitative RT-PCR analysis of the metabolic enzymes in the CD8 T cells cultured under as in the indicated conditions for 36 h. The results are presented relative to the expression of Cd3ε mRNA with the standard deviation (n = 3: technical replicates). f Naive CD8 T cells were stimulated with anti-TCR-β, anti-CD28 mAbs plus IL-2, and DM-α-KG without glutamine in the presence or absence of rapamycin (10 nM) for 36 h, and then the ECAR (left) and the OCR (right) were determined. Glycolysis was calculated by subtracting the ECAR before glucose injection from the ECAR 20 min after injection. Basal mitochondria respiration was calculated by subtracting the OCR in the presence of rotenone/antimycin A from the basal OCR at 15 min after the start of measurement. g Menin prevents effector CD8 T-cell dysfunction by targeting mTORC1-dependent metabolic activation. e, f *p < 0.05, **p < 0.01 (Student’s t-test)
