For blood analysis, freshly prepared plasmas were diluted twice before analysis. For tissue analysis, freshly sampled WAT and BAT were washed in PBS, weighed and incubated in free DMEM (Dulbecco’s modified Eagle medium) for 2 hours at 37°C. Media were kept for various analyses of secreted proteins.
Leptin and cytokines were assayed using “Mouse Leptin Kit” (Meso Scale Discovery, # K152BYC) and “mouse V-PLEX Proinflammatory Panel 1 Kit” (Meso Scale Discovery, # K15048D) respectively, on a QuickPlex SQ 120 apparatus (Meso Scale Discovery) following manufacturer’s instructions. IL-1RA levels were assayed using mouse IL-1RA Elisa kit (#EMIL1RN) from Thermo Fisher Scientific (Courtaboeuf, France). Glycerol and triglycerides determinations were performed using dedicated kit (Free Glycerol reagent and Triglyceride reagent, Sigma Aldrich).
Leptin and cytokines were assayed using “Mouse Leptin Kit” (Meso Scale Discovery, # K152BYC) and “mouse V-PLEX Proinflammatory Panel 1 Kit” (Meso Scale Discovery, # K15048D) respectively, on a QuickPlex SQ 120 apparatus (Meso Scale Discovery) following manufacturer’s instructions. IL-1RA levels were assayed using mouse IL-1RA Elisa kit (#EMIL1RN) from Thermo Fisher Scientific (Courtaboeuf, France). Glycerol and triglycerides determinations were performed using dedicated kit (Free Glycerol reagent and Triglyceride reagent, Sigma Aldrich).