D03021303

All procedures were approved by the Duke University Institutional Animal Care and Use Committee. Mice had free access to food and water and were fed a standard chow (Purina Rodent Chow no. 5001, Purina Mills, St. Louis, MO, USA) prior to being fed a semi-purified 10% fat diet (Research Diets, D12450B) or a 45% fat diet (Research Diets, D03021303) for 12-weeks. The genetic strategy used for generating CrAT^fl/fl and CrAT^skm−/− mice was described previously (Seiler et al., 2015 (link)). Glucose tolerance and insulin tolerance tests were performed in the morning 1 hour after food withdrawal. Mice were singly housed with access to water (gel pack). After taking glucose reading using a glucometer at the 0 minute time point, mice were given an intraperitoneal injection of 2.0 g glucose per Kg body or 0.08 U/mouse Humulin, both dissolved in saline. Blood for ELISA insulin measurements was collected at 0, 15, and 60 min. At the time of harvest for proteomics analysis, weights of mice fed the high fat diet averaged 38.6 g (1.1 STE, n=5) for CrAT^fl/fl and 41.1g (1.8 STE, n=8) for CrAT^skm−/−. At time of the glucose tolerance test, animals weighed 27.5 g (STE 0.8, n=5) for CrAT^fl/fl and 37.5g (1.6 STE, n=8) for CrAT^skm−/−.