Fig. 7.

ATP production rate of zebrafish liver cells exposed to 100 nM phoenixin-20 (PNX-20) during 1.5 h. ATP production rate was measured using the Seahorse XFp Real-Time ATP Rate assay. This assay is based on the kinetic quantification of oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). OCR and ECAR rates are first measured in basal conditions. Next, injection of oligomycin results in an inhibition of mitochondrial ATP synthesis that results in a decrease in OCR, allowing mitochondrial ATP production rates to be quantified. Complete inhibition of mitochondrial respiration with rotenone + antimycin A enables calculation of mitochondrial-associated acidification, allowing calculation of glycolytic ATP production. Data are presented as means ± SE (★P < 0.05, n = 3 wells/group). Student’s t test was used for statistical analysis.