Figure 1.

Metabolic phenotyping and effects of instantaneous IA additions on mitochondrial and glycolytic activities in MCF-7 and MDA-MB-231 cells. (A,B) MCF7 or MDA-MB-231 cells were cultured in XF24-well cell culture microplates (Seahorse Bioscience) at a density of 4 × 10⁴ cells/per well and then incubated for 24 h at 37 °C under 5% CO₂ atmosphere in proper media. OCR (A) and ECAR (B) values were normalized to total cell numbers for each cell line in the assay. Data are representative of at least three independent experiments, each with 3–5 statistical replicates and error bars represent SEM. (C) Relative ECAR and OCR data from (A) and (B) were plotted simultaneously to reveal overall relative basal metabolic profiles for each cell model. (D,E) Instantaneous effects of four successive infusions of 5-µM-IA on basal mitochondrial respiration in MCF7 (D) or MDA-MB-231 (E) cells. (F,G) Instantaneous effects of four successive infusions of 5-µM-IA on basal glycolytic activities in MCF7 (F) or MDA-MB-231 (G) cells. (H) Effects of increasing concentration of instantaneously added IA on both OCR and ECAR measures in both cell types.