Insulin Tolerance Test (ITT)

An insulin tolerance test (ITT) was performed after 6 weeks of treatment (W12) to allow for a 2-week rest period between metabolic tests. The ITT was also performed according to Beguinot & Nigro with the aforementioned fasting modification (24). During the ITT, mice were fasted for 6h prior to baseline blood glucose measurements and intraperitoneal injection of 0.5IU/kg insulin (Humulin^® R U-100, Lilly, Cat. No. HI-210) diluted in 0.9% NaCl in an injection volume of 3.6µL/g body weight. Thereafter, blood glucose measurements were taken at 15, 30, 60, and 90-minute timepoints. If blood glucose levels dropped below 36mg/dL, mice were rescued via injection of 20% aqueous glucose solution (Sigma, Cat. No. G8270).