Am1241

Following the acclimatization period, rats received a high-fat diet (HFD) 21% fat content (equating to 40% digestible energy) from lipids (Specialty Feeds SF00-219, Glen Forrest, WA, Australia) for 9 weeks, as described in our previously published study [13 (link)]. Throughout the study, animals could access food and water ad libitum. Animals were then maintained on the HFD and treated for a further six weeks with a daily i.p. injection, with either vehicle (0.9% isotonic saline solution containing 0.75% Tween 80: n = 9–10), 3 mg/kg body weight of AM1241 (Cayman Chemicals, Ann Arbour, MI, USA), or 0.3 mg/kg body weight of AM630 (Cayman Chemicals, Ann Arbour, MI, USA) dissolved in the vehicle solution (n = 9–10). These compounds and their doses were chosen at the time of the initiation of the study due to the following papers: AM1241 [32 (link)] and AM630 [31 (link)]. EchoMRI Whole-Body Composition Analyzer (EchoMRI-900; EchoMRI, Houston, TX, USA) was used to determine body composition, as previously described [6 (link)].
Following treatment, rats were anesthetized with 3% isoflurane inhalation (Abbott Laboratories, Chicago, IL, USA) with cardiac blood collected to confirm their death; then, all other major organs including fat pads were removed post-mortem, weighed, snap frozen in liquid nitrogen, and stored at −80 °C for further analyses.