Extracellular metabolic flux analysis

XH Xia He
GC Guangfan Chi
ML Meiying Li
JX Jinying Xu
LZ Lihong Zhang
YS Yaolin Song
LW Lina Wang
YL Yulin Li
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We seeded MEFs and ciXEN cells onto Matrigel-coated XF96 well plates (Seahorse Bioscience, USA). To obtain extracellular acidification rate (ECAR), the cells were metabolically perturbed by glucose (10 mM), oligomycin (1 mM), and 2-deoxyglucose (50 mM) (all from Sigma). For mitochondria stress tests, oxygen consumption rate (OCR) was measured by supplementing with DMEM, 1 μM oligomycin, 2 μM FCCP, and 1 μM rotenone and 1 μM antimycin A (all from Sigma). The final measurements were normalised by the number of viable cells.

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