Male aged-matched C57/Bl6 mice (Jackson Laboratories) were placed on 60% HFD (Research Diets, Cat# D12492) at 6 weeks of age or maintained on chow (Teklad Diets, Cat# 8904) for 12 weeks. After 12 weeks, mice were either maintained on chow or HFD or switched to chow for the remaining 12 weeks (Figure 1a). Animals were grouped, housed and maintained on a 12-h light-dark cycle in pathogen-free facilities at the Ohio State University. At the end of the diet, mice underwent insulin tolerance testing (ITT) and intraperitoneal glucose tolerance testing (IpGTT), and measurement of body weight, adiposity and circulating levels of insulin and glucose. After 6 h fast (9 am–2 pm) for ITT or overnight fast (5 pm–9 am) for IpGTT, insulin (0.5 U/kg) or glucose (1 g/kg) was administered via intraperitoneal injection at T0 after obtaining fasting blood samples [14 ,15 (link)]. All experiments were approved by and in accordance with The Ohio State University Institutional Animal Care and Use Committee (2014A00000108-R1, approval date: 09/21/2017).