Research Paper Volume 14, Issue 23 pp 9466—9483

Figure 9. Treatment with glucosamine hydrochloride reduces mitochondrial oxygen consumption rates in MCF7 cells. Cells were seeded and treated with glucosamine, as described above. Briefly, cells were seeded at a density of fifteen thousand in a 96-well format. (A) Extracellular consumption rate (ECAR) was assessed by Seahorse metabolic flux analysis. A representative trace is shown in the top panel. Importantly, glucosamine treatment only had minor effects on glycolysis. (B) Oxygen consumption rate (OCR) was measured by Seahorse metabolic flux analysis. A representative trace, in the top panel, shows decreased OCR in samples treated with glucosamine (20 mM), versus the vehicle alone control cells. The bar graph (lower panel) shows that glucosamine treatment significantly decreases the basal respiration, ATP production, maximal and spare respiration, as compared to the control cells. In panels A and B, experiments were performed three times independently, with six repeats for each replicate. Bar graphs are shown as the mean ± SEM, t-test, two-tailed test. *p < 0.05, ***p < 0.001.