Figure 5 from Multistate Gene Cluster Switches Determine the Adaptive Mitochondrial and Metabolic Landscape of Breast Cancer

Metabolic wiring of the switch positions probed by ¹³C-labeled substrates. A, Nutrient supply and ¹³C-labeling conditions. High and low glutamine (H/L = 1/0.1 mmol/L) levels were combined with the presence and absence of pyruvate (+/−, 1/0 mmol/L), and glucose was kept at 10 mmol/L. B, Glucose uptake, lactate secretion, and fractional enrichment of m + 3 labeled intracellular lactate. Cells were grouped by switch state. Two-way ANOVA with Sidak multiple comparisons test. C, Fractional enrichment of the m+3 pyruvate isotopomer, following U13C-glucose labeling for 24 hours, in cells grouped by switch state. Two-way ANOVA with Sidak multiple comparison tests. D, Overall model of the MB1_UF/MB1_LF metabolic switch in central carbon metabolism. E, Relative carbon contribution to citrate following 24 hours labeling with U13C-glucose and U13C-glutamine, in cells grouped by switch state. Two-way ANOVA with Sidak multiple comparison tests. F, Fractional enrichment of the m+2 (PDH route; left) and m+5 (PDH + PC route; right) isotopologues in citrate following 24 hours labeling with U13C-glucose, in cells grouped by switch state. Two-way ANOVA with uncorrected Fisher LSD multiple comparison tests. G, Glutamine uptake, glutamate secretion, and fractional enrichment of malate m+4 (oxidative route) and citrate m+5 (reductive carboxylation) isotopologues following 24 hours labeling with U13C- glutamine (right panels). Two-way ANOVA with uncorrected Fisher LSD multiple comparison tests. H, Substrate dependence of basal oxygen consumption rate and total cellular ATP content in cells grouped by switch state. Two-way ANOVA with uncorrected Fisher LSD multiple comparison tests. ns, nonsignificant, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.00; ****, P ≤ 0.0001.