Methionine depletion induces a lower metabolic state and cell cycle block in ML cells. (A-C) The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) of AML cell lines (n = 3 biological replicates), PBMC-derived CD34⁺ cells (n = 1), as well as a patient AML sample (AML#17), as measured by Seahorse bioassay. (D-E) Forty-eight-hour glucose consumption and lactate production per cell per hour as determined enzymatic spectrophotometric assays (n = 3). (F) Tetramethylrhodamine, methyl ester (TMRM) mean fluorescent intensity (MFI) determined mitochondrial membrane potential (n = 3). (G) 1D 1H−nuclear magnetic resonance (NMR) spectroscopic metabolite intensity fold changes, control normalized to 1 (3 biological replicates measured once). (H) Hoechst-Pyronin Y cell cycle staining and Annexin-V PI staining of apoptosis of MOLM13 cells (representative plots of n = 3 experiments) and PBMC-derived CD34⁺ (n = 1) cells depleted of methionine for 24 hours. All experiments: error bars represent mean ± SEM. Statistical analysis by Student t test, ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001.