Oxygen consumption rate (OCR)
Oxygen consumption rate (OCR) was measured using the XFe96 analyzer
(Agilent Technologies). Briefly, cells (4 X 104 cells)
were plated into wells of an XFe96 cell culture microplate and incubated
at 37 ⁰C in a CO2 incubator for 24 hours. The assay was
started after cells were equilibrated for 1 hour in XF assay medium
supplemented with 10 mM glucose, 1 mM sodium pyruvate and 2 mM
glutamine. OCR activity was monitored through sequential injections of 1
µM oligomycin, 0.6 μM FCCP, and 1 μM rotenone/ antimycin A to calculate
basal, coupled, maximal respiration and spare respiratory capacity. Each
plotted value was normalized either to cell counts. Agilent Seahorse XF
Analyzer measures oxygen consumption rate (OCR) at intervals of
approximately 5-8 min. OCR data were evaluated using 8 individual well
values.