Citrate synthetase Antibody - #DF13222

FIGURE 4 SYSU-3d promotes lipid oxidation by activating the heat shock factor 1 (HSF1)/ PPARγ coactivator-1α (PGC-1ɑ) metabolic pathway (a–e) HepG-2 cells were exposed to palmitic acid (PA; 0.75 mM) in the presence or absence of SYSU-3d (1 μM) for 24 h and then analysed. (a) mtDNA/nDNA. (b) Determination of the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) from metabolic flux analyses. (c) Free fatty acid (FFA) uptake assay using the probe TF2-C12. (d) ROS level determination by flow cytometry. (e) Expression of citrate synthase (CS), malate dehydrogenase 2 (MDH2) and mitochondrial complexes; protein levels were quantified and normalized to Actin. (f–h) Determination of CS, mitochondrial complex I and II activity by normalizing to protein level. (i–k) Determination of oxygen consumption rate (OCR) ability, mitochondrial copy numbers (indicated by Mito-tracker probe) and ROS level (indicated by DCFH-DA probe) in HSF1−/− cells. (l and m) Determination of triglyceride (TG) levels, cell survival, mitochondrial copy numbers and ROS levels in PGC-1ɑ−/− cells. (n) Determination of OCR in PGC-1ɑ−/− cells. (o) Expression of metabolic regulators and inflammation- and apoptosis-related proteins in PGC-1ɑ−/− cells; protein levels were quantified and normalized to Actin; n = 5 independent experiments. *P