C57bl 6j

Specific pathogen-free C57BL/6J and Nos3^-/- (C57BL/6J genetic background) mice were purchased from Orient Bio (Gyeonggi, South Korea) and from The Jackson Laboratory (Bar Harbor, ME, USA) and bred in our pathogen-free animal facility. Animal care and all experimental procedures were conducted in accordance with the approval and guidelines of the INHA Institutional Animal Care and Use Committee (INHA IACUC) of the Medical School of Inha University. Control animals were fed a normal diet and experimental animals were fed a diet containing 4% cholesterol and 1% cholic acid (Feed Lab. Co., Gyeonggi, South Korea) for 3 months. Treatments of mice were performed as described in the Supplemental information online.

The Ajou University Animal Care and Use Committee approved all animal studies (IACUC2015-0001), and the experiment conformed to the Guide for the Care and Use of Laboratory Animals published by the United States National Institutes of Health. All experiments were performed in accordance with relevant guidelines and regulations. Six-week-old C57BLKS/J-Lepr^db/Lepr^db or wild type C57BL/6J male mice were purchased from Orient Bio, Inc. (Seongnam, Korea) and acclimatised for one week. Mice were randomly grouped and orally administered with vehicle, WY14643 (20 mg/kg), rosiglitazone (20 mg/kg), or MD001 (5 mg/kg or 20 mg/kg) once a day for two months. For the oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (IPITT), mice were fasted for 12 h and were treated with sterile glucose (1 g/kg, Sigma-Aldrich/Millipore) or human insulin (1 unit/kg, Eli Lilly and Company, Indianapolis, IN, USA). Blood glucose levels were measured at the indicated time point using an OneTouch Ultra Blood Glucose Monitoring System (LifeScan, Inc., Milpitas, CA, USA). For the toxicity study, wild type C57BL/6J male mice were randomly grouped and orally administered with vehicle and MD001 (50 mg/kg or 100 mg/kg) once a day for two month. Blood cells, body weight change, and blood and urine metabolites were analysed for toxicity.

C57BL/6J and C57BL/6N male mice were obtained from OrientBio (Kyounggido, Korea). Mice were housed at the animal facility of the Lee Gil Ya Cancer and Diabetes Institute (CACU, Gachon University, Incheon, Korea), in accordance with “Guidelines and Animal for Users”. Six week-old male C57BL/6J mice were fed an HFD (60% fat primarily from lard, Research Diets, Inc., New Brunswick, NJ, USA, #D12492). After 10 weeks (at 16 weeks of age), mice were randomly divided into two groups: the PBS-treated HFD group (HFD + PBS; n = 5) and the phloroglucinol-treated HFD group (HFD + phloroglucinol; n = 5). Phloroglucinol (100 mg/kg in PBS) or PBS was administered by oral intubation daily for 9 weeks. Body weight and food consumption were measured weekly.