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Figure 2. The metabolic phenotype of the highly metastatic MDA-MB-231HM.LNm5 (MDA-231HM.LNm5) and parental MDA-MB-231 (MDA-231) cell lines. Baseline oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) calculated in the presence of glucose were combined to generate the basal phenogram (A), while maximum ECAR (Oligomycin treated) and maximum OCR [carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) treated] were combined to generate the maximum phenogram (B). The glycolytic profile shows ECAR readings in the presence of glucose (11 mmol/L) from which the basal glycolytic rate was calculated (C) [Supplementary Figure 1]. Following injection of oligomycin (5 μmol/L), both maximum glycolytic rate (G) and glycolytic reserve (H) could be determined. The respiration profiles of both cell lines (D) show OCR readings in the presence of glucose and following subsequent addition of oligomycin (5 μmol/L), FCCP (1 μmol/L) and finally antimycin plus rotenone (2.5 μmol/L of each). This procedure allows the quantification of basal mitochondrial respiration (E) and ATP production (F), as well as maximal mitochondrial respiration, spare respiration capacity, proton leak and non-mitochondrial respiration [Supplementary Figure 2]. Data are presented as mean ± SEM, n = 5-7. The student’s t-test was used to test for statistical significance. NS: not significant; *P < 0.05, **P < 0.01