Sunflower oil

Clozapine n-oxide (Tocris, Bristol, UK) was dissolved at 1 mg ml⁻¹ in sterile saline. Salvinorin B (SalB; Cayman Chemical, Ann Arbor, MI, USA) was suspended in DMSO (25 mg ml⁻¹) and then diluted to 5 mg ml⁻¹ in sunflower oil (Sigma, St Louis, MO, USA). 4-OH-Tamoxifen (Sigma) was dissolved in DMSO (20 mg ml⁻¹) and then diluted to 8 mg ml⁻¹ in sunflower oil (Sigma). R-amino-methylhistamine (RAMH, Tocris), Immepip (Tocris) and JNJ JNJ5207852 (Tocris) were dissolved at 9 mg ml⁻¹ in sterile saline. All drugs were diluted such that mice received 0.05 ml per 10 g for each drug, for all experiments.

All animal experiments were approved by the Institutional Animal Care and Use Committee of Asan Institute for Life Sciences (Approval No. 2020-12-131), and studies were conducted in accordance with the approved guidelines and regulations. LKB1^ΔIEC mice were generated by crossing LKB1^f/f and Villin^Cre mice (Jackson Laboratory, Bar Harbor, ME). LKB1^ΔIEC mice were co-housed with their control littermates (LKB1^f/f) after genotyping. All mice were maintained in groups of 2 to 5 in cages under specific pathogen-free conditions and received sterilized food and water ad libitum. All experiments were performed with sex- and age-matched 6- to 12-week-old mice. Mice were fasted 4 hours before euthanasia, and blood and tissues were collected under anesthesia with a mixture of ketamine (100 mg/kg) and xylazine (20 mg/kg). For HFD experiments, LKB1^f/f and LKB1^ΔIEC mice were fed a HFD (60% of total calories from fat) for 8 weeks. OCA (AdipoGen, San Diego, CA), dissolved in 10% DMSO (BioLife Solution, Bothell, WA) and 90% sunflower oil (Sigma Aldrich, St. Louis, MO), was administered by oral gavage at a dose of 30 mg/kg for 5 days. Vitamin A (retinyl palmitate, Sigma Aldrich, 100 mg/kg) in sunflower oil was administered by oral gavage at the beginning and end of the dark cycle (16 and 4 hours before death), described previously.³⁸ (link)