Extended Data Fig. 6: The effect of co-inhibiting glutaminases and amidotransferases on metabolism and tumour cell proliferation.
From: Identifying strategies to target the metabolic flexibility of tumours
a–c, The effect of DON (50 mg/kg, 4 h) on either CT or Gls1KO/shGls2 tumours from animals treated prior to [U-13C]glutamine bolus: . a, Total concentration of glutamine-derived amino acids (n = 3 mice per group); b, Total concentration of Krebs cycle intermediates (n = 3 mice per group); c, 2D 1H-13C-HSQC (heteronuclear single quantum coherence spectroscopy) NMR signals of glutamine and glutamate. d, Enrichment from either [U-13C]glutamine or [U-13C]glucose in HCCMYC-CT and HCCMYC-Gls1KO/shGls2 tumour cells treated with DON for 3 h (n = 3 independent experiments). GC-MS. e, Isotopologue distribution of the 13C incorporation into malate in the experiment shown in (d) (n = 3 independent experiments). Data are presented as mean ± S.D. Statistical analysis was performed using a two-tailed Student’s t-test. Complete list of exact P values is provided as a source data file. f, Combination of glutaminase inhibition and DON on cell proliferation of HCCMYC cells: HCCMYC-CT cells were treated with 1 μM CB-839 and/or 2 μM of DON. g, h, HCCMYC-Gls1KO/shGls2 (g) and HCCMYC-CT (h) cells treated with a combination of DON and CB-839 with or without the addition of the indicated amino acids. AAAP - a mix of alanine, aspartate, asparagine and proline. i, HepG2 cells treated with DON and/or CB-839 at the indicated concentrations. In (f–i) growth was monitored in an IncuCyte Live-Cell analysis system. In (f–i), representative curves from one of three independent experiments with 3 replicates are shown. Data are presented as mean ± S.D.
