Glutamine controls mitochondrial OXPHOS in AML. (A) A total of 11 AML cell lines were cultured with or without Gln (4 mM) for 48 hours. Apoptosis was quantified by flow cytometric analysis of Annexin V binding. (B) OCI-AML2 and MOLM-14 cell lines were cultured for 48 hours with or without Gln (4 mM) and αKG (5 mM) as indicated, and apoptosis was quantified based on Annexin V binding. (C) OCI-AML2 and MOLM-14 cell lines were cultured for 48 hours with or without Gln (4 mM), and the OCR was measured using a Seahorse XF96 extracellular flux analyzer, under both basal conditions and after the addition of increasing doses of CCCP (0.25, 0.5, and 0.5 µM) and antimycine (10 µM), as indicated. Histograms show data that are representative of 3 independent experiments at baseline or after CCCP (cumulative dose, 1 µM). (D) MOLM-14 cell lines were cultured for 48 hours with or without Gln (4 mM) and αKG (5 mM) as indicated, and the OCR was measured as in C. Histograms show data that are representative of 3 independent experiments. *P < .05, **P < .01, ***P < .001.