H10060

Male C57BL/6J mice and ob/ob mice were obtained from the Beijing Huafukang Bioscience (Beijing, China). Gfral-knockout mice were purchased from Cyagen Biosciences (Guangzhou, China). Animals were housed in the Northwest A&F University animal facility under standard conditions with 12 h of light and 12 h dark cycle with free access to food and water. Animals were randomly divided into different groups as specified. The experimental HFD diet (60% calories from fat, 20% calories from protein, and 20% calories from carbohydrate, H10060) was purchased from Beijing Huafukang Bioscience (Beijing, China).

The IACUC number for animal ethics approval is 2468. All animal studies were approved by the Institutional Animal Care and Use Committee of Tongji Medicine College. All applicable institutional guidelines for the care and use of animals were followed. Six-week-old female C57BL/6J mice were purchased from Beijing HFK Bioscience Co., Ltd., and raised in specific pathogen-free (SPF) conditions in the Tongji Medicine School Animal Center. The mice were housed at 22 ± 2°C on a 12-h light cycle. For the diet-induced obesity (DIO) model, the mice were fed a high-fat diet (HFD) (H10060, 60% fat, 20% protein, 20% carbohydrate, Beijing HFK Bioscience Co., Ltd.) for 10 weeks to induce obesity before SG surgery (17 (link)).

WT and BAP31-ASKO mice (6-week-old) were fed with a high-fat diet (HFD. H10060, Beijing HFK Bioscience Co. Ltd.) for 14 weeks. Body weight (BW) was measured every 4 days. After exposure for 13 weeks, mice were food deprived for 16 or 5 hours, and then injected with glucose (2 mg/kg) or insulin (0.75 U/kg) solution intraperitoneally. The blood glucose at 0, 15, 30, 60, and 120 minutes were determined using a one-touch glucometer (Bioland technology, Shenzhen, China).

The experimental subjects of this study were 24 SPF-grade C57BL/6J male mice (SYXK[Jun] 2015-0004, Hebei In vivo Biotechnology Co., Ltd), 6 weeks old and weighing 17.8–21.6 g. The mice were housed in the Animal Experiment Center of Hebei General Hospital, with 4 mice per cage, and maintained at 20–25 °C, 50–55% humidity, with artificial light source simulating a circadian cycle (12 h/12 h). Eight healthy mice were randomly assigned to the normal control group (group C) and fed with regular chow (D1035, 10% energy from lipids, Beijing Huafukang Biotechnology Co., LTD, China), free food and water. The rest were fed with high-fat diet (H10060, 60% energy from lipids, Beijing Huafukang Biotechnology Co., LTD, China) for 12 weeks. An average weight increase of 20% compared with the normal-diet group was used as the criterion for obese mice. The obese mice were randomly divided into the model group (high-fat diet, group H) and semaglutide group (H+ semaglutide, group S), comprising 8 mice in each group. Mice in group S were treated with semaglutide 30 nmol/kg/d for 12 weeks whereas mice in groups H and C were treated with equal amounts of saline. The study followed the guidelines of the Guide for the Care and Use of Laboratory Animals and was approved by our Animal Ethics Committee (No. 202173). All methods below are from previous studies [7 (link)].

Six‐week‐old male C57BL/6N mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd, and housed in a controlled condition (22 ± 2°C, 40%–60% humidity, and 12 h light–dark cycle). After 1 week of adaptation, the mice were randomly divided into control group (Con), HFD group, and MP group (n = 8–10). The mice in the Con group were fed with standard diet (13.5% of energy from fat; 1022; Beijing HFK Bioscience Co. Ltd) and other mice were fed with a HFD (60% of energy from fat; H10060; Beijing HFK Bioscience Co. Ltd). Mice in the MP groups were, respectively, gavaged with high dose of MP solution (HMP‐100 mg kg⁻¹ day⁻¹) and low dose of MP solution (LMP‐50 mg kg⁻¹ day⁻¹), and other mice were administrated with water as control. After 20 weeks, the animals were subjected to the metabolic cages, and food intake data were collected over 24 h. Feces were collected for 16S sequencing analysis. The animals were fasted for 10 h prior to euthanasia, and blood samples and vital organs were collected. This study was approved by the Laboratory Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM‐LAEC2020049).

After 3 days of adaptive feeding, mice were randomly divided into 2 groups, CD and HFD groups. Mice in CD and HFD groups were fed a control standard diet [20% kcal protein, 70% kcal carbohydrate and 10% kcal fat, H10010, Beijing HFK BIOSCIENCE CO.,LTD.) or high-fat diet (20% kcal protein, 20% kcal carbohydrate and 60% kcal fat, H10060, Beijing HFK BIOSCIENCE CO.,LTD.)], respectively. Body weights were measured once a week. After 8 weeks of feeding, mice were randomly selected from the CD group and HDF group for examination via glucose tolerance test (GTT), insulin tolerance test (ITT), fasting blood glucose (FBG), HOMA-IR, and serum insulin levels to establish the model.