Xf cell q17 mito stress test kit

OCRs were measured using the XFe96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA, USA). Cardiomyocytes were seeded (10 000 cells/well) and incubated overnight in DMEM supplemented with 10% (v/v⁻¹) FBS. One hour before the measurement, cells were incubated with XF medium at 37 °C in a CO₂-free incubator. The OCRs were detected under basal conditions and after the application of 1 μM oligomycin, 0.5 μM FCCP and 0.5 μM rotenone+0.5 μM antimycin A (XF Cell Q17 Mito Stress Test Kit; Seahorse Bioscience). Each sample was analyzed four times.
Extracellular acidification rates (ECRs) were detected under basal conditions and after the application of 10 mM glucose, 1 μM oligomycin and 50 mM 2-DG (XF Cell Q17 Glycolysis stress test kit; Seahorse Bioscience). Each sample was analyzed four times.

OCR measurement was done using the XFe96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA, USA). Differentiated adipocytes were seeded (10⁴ cells/well) and incubated overnight in DMEM supplemented with 10% FBS. The cells were treated with Rg5 (10 μM) or metformin (1 mM) for 4 h. One hour before the measurement, the cells were incubated with XF medium at 37°C in a CO₂-free incubator. The OCR was detected under basal condition and after the application of 1 μM oligomycin, 0.5 μM FCCP (carbonyl cyanide p-trifluoromethoxyphenylhydrazone) and 0.5 μM rotenone + 0.5 μM antimycin A (XF Cell Q17 Mito Stress Test Kit; Seahorse Bioscience). OCR result was analyzed using the Seahorse XF96 software. Each sample was analyzed four times.