Figure 6.

Glycolytic and respiratory capacity and nutrient usage in RT112 and 5637 cells. (a) PER and OCR values (attributable to glycolysis and oxidative phosphorylation respectively) in basal condition, maximal capacity (reached with treatment with optimized concentration of oligomycin A for maximal glycolytic capacity or FCCP for maximal respiratory capacity) and under 5 h-glucose deprivation. Seahorse data were normalized on cell number measured after Hoechst 33342 staining as blue-positive objects acquired by Operetta CLS™ and counted using Harmony software. (b) The ability to oxidize three different nutrients (glucose, glutamine and fatty acids) reported as dependency and flexibility from XF Mito Fuel Flex Test performed using Agilent Seahorse XFe96 Analyzer. (c) OCR values measured in presence/absence of 5 µM Etomoxir from XF Palmitate-BSA FAO Substrate assay using Agilent Seahorse XF24 Analyzer. OCR values were normalized on protein content measured by Bradford assay. All results are the mean of at least three experimental replicates. Statistical test: t-test, ** for p < 0.01.