Effects of the GLS inhibitor CB-839 on BM cells in vitro. (A) Experimental scheme. Seahorse metabolic flux assay was performed with BM cells from WT mice (n = 6) or VF mice (n = 6). The cells were treated with DMSO or CB-839 (10 μM) 1 hour before the Seahorse assay. (B) Analysis of changes in OCRs upon administration of oligomycin, FCCP, and Antimycin A (left); maximal OCR values (middle) and maximal OCR displayed by sorted CFU-E (lin− ckit+ Sca1− CD41− CD16− CD150− CD105+) (right). (C) Analysis of changes in ECAR (mpH/min), indicative of glycolysis, upon administration of glucose, oligomycin, and 2-DG (left); maximal glycolysis rate (middle) and maximal glycolysis rate from sorted CFU-E cells (right), gated as described in panel B. The plots represent the mean ± standard error of the mean (SEM) from 6 mice per group from 1 experiment (for CFU-E panel, n = 1). 2-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison was performed. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone.