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FASEB J (2015) 29(7):302739

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Oxygen consumption assays Primary brown adipocytes were seeded into 24-well V28 plates (22,000 cells per well), differentiated as described above and Cell Mito Stress Tests were performed in a Seahorse XFe24 as previously described 74 In vitro lipolysis and lactate assays Lipolysis in differentiated adipocytes was assessed after overnight serum starvation, with or without addition of 500 ng/ml vaspin for 30 min prior measurements under basal conditions and after addition of CL, FSK or ISO (final concentrations 1 M or 100 nM) for 12 h using standard assays according to the manufacturers protocol (assay medium 2% free fatty acid free BSA in DMEM

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