Fig 1: P2RX4/SLC25A11 mediates the intracellular transport of AKG.(A and B) Intracellular and medium 13C-AKG level (A) and intracellular total AKG level (B) (n = 3). (C and D) Cytosol (mitochondrial-free) (C) and mitochondrial (D) AKG level (n = 6). (E) Relative changes of dicarboxylate carriers in response to AKG treatment (n = 3). (F) SLC25A11 protein expression of mitochondrial membrane protein in AKG-treated primary hepatocytes (PA treatment) (n = 6). (G) Immunoblots of SLC25A11 in si-SLC25A11–treated primary hepatocyte (n = 3). (H to J) Immunoblots (H) and quantification (I), and enzyme activity (J) of PEPCK, G6Pase, and FBP in primary hepatocyte (PA treatment) (n = 4 to 8). (K) Relative changes of purine receptors in response to AKG treatment (n = 3). (L) Schematic representation of Biacore. (M) Immunoblots of protein expression after AKG pull-down in mice liver. (N) Immunoblots and quantification of P2RX4 in the liver. Twelve-week-old male mice were treated with vehicle or 5-BDBD (P2RX4 antagonist, 5 mg/kg) (n = 3). (O) Cytosol (mitochondrial-free) and mitochondrial AKG level (n = 6 per group). (P) Immunoblots and quantification of P2RX4 in si-P2RX4–treated primary hepatocyte (n = 3). (Q to S) Immunoblots (Q) and quantification (R), and enzyme activity (S) of PEPCK, G6Pase, and FBP in si-P2RX4–treated primary hepatocyte (PA treatment) (n = 4 to 8). (T) Graphic abstract mechanism of AKG on gluconeogenesis. Results are presented as means ± SEM. (B, F, G, N, and P) *P = 0.05 by nonpaired Student’s t test. In (C), (D), (I), (J), (O), (R), and (S), different letters between bars indicate P = 0.05 by one-way ANOVA followed by post hoc Tukey’s tests.