Fig. 2.

Pyruvate impairs sensitivity to glutaminase inhibition by increasing TCA cycle anaplerosis. Increasing sodium pyruvate concentration in RPMI 1640 + 5% FBS increases CB-839 IC₅₀ (mean ± SEM, n = 3–7, one-way ANOVA) in a MDA-MB-231, b BT549 or c Hs578T cells during a 3 day assay. 24 h treatment with CB-839 decreased fumarate level in MDA-MB-231 cells in RPMI 1640 + 5% FBS but not RPMI 1640 + 5% FBS supplemented with 1 mM sodium pyruvate. Fumarate level was normalised to either initial cell number seeded (d) or protein amount at endpoint (e) (mean ± SEM, n = 3–4, two-way ANOVA)