Assaying glutamine oxidation and demonstrating transaminase pathway activity.

<p>A. Schematic illustration of biochemical pathway for glutamine oxidation in the mitochondria. B. Kinetic OCR response in SF188f cells to glutamine (4 mM). SF188f cells were plated at 20,000 cells/well in XF24 cell culture plates 24–28 hours prior to the assays. The assay medium was the substrate-free base medium. The OCR value was not normalized. A representative experiment out of three is shown here. Each data point represents mean ± SD, n = 4. C. OCR response (% of baseline) in SF188f cells to glutamine (4 mM) and AOA (100 µM). Glutamine-induced OCR reached 60% over the baseline (OCR at measurement 6 divided by that at measurement 3) while AOA addition reduced it to 20% (OCR at Measurement 9 divided by measurement 3). SF188f cells were plated at 20,000 cells/well in XF24 V7 cell culture plates 24–28 hours before the assays. The % OCR was plotted using measurement 3 as the baseline. The assay medium was the substrate-free base medium. A representative experiment out of three is shown here. Each data point represents mean ± SD, n = 4.