PMC:6879884 / 10496-11605 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/6879884","sourcedb":"PMC","sourceid":"6879884","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6879884","text":"Seahorse glycolysis stress assay.\nHuman bronchiolar epithelial cells were seeded into a Seahorse XF96 plate and incubated at 37°C, 5% CO2 for 48 h. The medium was changed 24 h before the Seahorse experiment, and cells were exposed to 5 or 15 mM glucose with or without BrPy (100 μM) or epalrestat (1 or 10 μM) for the last 30 min before the Seahorse glycolysis stress assay was performed according to the manufacturer’s instructions followed by the sequential injection of oligomycin to inhibit ATP-linked reparation and 2-deoxy-d-glucose (2-DG) to inhibit glucose metabolism. The plate layout was separated into quadrants to reduce edge effects. Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured. Glycolysis rate was calculated by subtracting the normalized ECAR values after 2-DG injection from the ECAR values after glucose injection to exclude the nonglycolytic acidification from the calculation. Glycolytic capacity was calculated by subtracting the nonglycolytic acidification rate (ECAR after 2-DG injection) from the maximum ECAR after 1 μM oligomycin injection.","divisions":[{"label":"title","span":{"begin":0,"end":33}}],"tracks":[]}