Animals
All animal experiment procedures were in compliance with the Animal Protection Act of Korea and the Guide for the Care and Use of Laboratory Animals. All studies were conducted at Biotoxtech Co., Ltd., Republic of Korea (Institutional Animal Care and Use Committees (IACUC) Approval No.: 150289) except for the metabolic assay and western blot assay, which were conducted at Asan Medical Center, Korea (approval from the Institutional Animal Care and Use Committee of Asan Institute for Life Sciences). Male C57BL/6J mice of (Jackson Laboratory, Maine, USA) 14 weeks of age (supplier: ORIENTBIO Inc., Republic of Korea) – fed HFD starting at 6 weeks of age – were purchased and housed in a controlled environment (12h light/dark cycle, light on at 7AM, 22 ± 2°C, humidity 54.4% ± 8%). After acclimation period of two weeks and randomized grouping, each animal was identified by individual color-coded cage card displaying group and dose level. Mice had ad libitum access to tap water and either pelleted rodent chow containing 10% fat diet (normal control) or 60% fat diet (negative control, positive control, and HSG4112 groups) (Research Diets Inc., USA). Individual food and drinking water consumptions were measured weekly for all groups unless noted otherwise. Animals were randomly separated into groups. Body weight was measured weekly, at minimum. Animals were dosed via gastric intubation with disposable syringes (1mL, Doowon meditec Co., Republic of Korea) fitted with an intubation tube once a day in the afternoon (3–4 PM), unless otherwise noted. All animals were fasted for 14-16 hours and anesthetized by inhalation of isoflurane before terminal sacrifice.