Fig. 5
Metabolic alterations in metastatic compared to nonmetastatic TNBC cells. a Relative basal oxidative consumption rate (OCR) of E-Wnt, M-Wnt, metM-Wntlung, as compared to M-Wnt cells. b Representative mitochondrial stress test of E-Wnt, M-Wnt, metM-Wntlung cells. Cells were injected with 1 μM oligomycin (oligo), followed by 1 μM carbonyl-cyanide-4-(trifluoromethoxy)phenyhydrazone (FCCP) and finally a combination of rotenone/antimycin (3 μM), (Rot/Ant). Uncoupling of the electron transport chain (ETC) from ATP synthesis (FCCP) reveals maximum oxidative respiration. Treatment with oligomycin allows the calculation of ATP production by the ATP synthase. c Representative glycolysis stress test showing extracellular acidification rate (ECAR) as a result of injection with 10 mM glucose, 1 μM oligomycin and 50 mM 2-deoxy-glucose (2DG). Response to glucose stimulation and relative maximal respiration are shown. All OCR and ECAR measurements were performed in sextuplicate and normalized to μg protein. Graphs show the mean are inclusive of at least three independent experiments. d Colony formation assay of E-Wnt, M-Wnt, metM-Wntlung cells grown in media containing 5 mM or 10 mM glucose or 10 mM galactose. Colonies were allowed to grow for 14 days before being fixed and stained. e Relative basal OCR and representative mitochondrial stress test of M6 and M6C cell lines. f Representative glycolysis stress test and relative glycolysis (ECAR) of M6 and M6C lines as compared with M6 cells. Graph shows mean ± SEM and is inclusive of three independent experiments. *p < 0.05, ***p < 0.001, Student’s t-test
