C57bl 6j wt mice

All animal experiments were approved by the Use and Care of Experimental Animals Committee of the Jichi Medical University Guide for Laboratory Animals, and were carried out in accordance with the Jichi Medical University guidelines. C57BL/6J WT mice were purchased from Japan SLC, Inc. (Hamamatsu, Japan). LMP7^−/− mice have been previously described4 (link). Male mice (9–10 week old) were fed with either 60 kcal% HFD (Research Diets: D12492, Japan LSG, Tokyo) or standard chow (CE-2; CLEA Japan Inc., Osaka). Each mouse was weighed every 2 weeks. At terminal points, mice were fasted for 6 h, weighed again and blood was collected to obtain serum. After perfusion, the tissues were carefully excised and weighed.

All animal experiments were conducted in accordance with the ethical guidelines of the Kyoto University animal experimentation committee and the Japanese Pharmacological Society. All animal use and study protocols were approved by the Kyoto University animal experimentation committee (approval number: 20-42). Male C57BL/6J WT mice (RRID: IMSR_JAX:000664) were purchased from Japan SLC, and TRPA1-KO mice were obtained from the Jackson Laboratory (Bar Harbor, ME; #006401). Mice were maintained in our laboratory and used at 8 to 12 weeks old and 20 to 30 g. TRPA1-KO mice were backcrossed to C57BL/6J mice for at least 10 generations and genotyped by genomic PCR using the following primers: 5′-tca tct ggg caa caa tgt cac ctg ct-3′ and 5′-tcc tgc aag ggt gat tgc gtt gtc ta-3′. All mice were housed at a constant ambient temperature of 22° ± 2°C under a 12-hour light/12-hour dark cycle and were allowed water ad libitum. The numbers of used animals were empirically determined and were similar to previous reports (9 (link), 29 (link), 47 (link)).

Animal experiments were performed as previously described^{12 (link)}. C57BL/6J WT mice or ob/ob mice at ten weeks of age were purchased from Japan SLC Inc. (Shizuoka, Japan). The phenotypic data of the mice are summarized in Table S1. Animal experiments were approved by the animal ethics committee of The University of Tokyo and according to the ARRIVE guidelines and the University of Tokyo guidelines for the care and use of laboratory animals. After purchasing, the mice were housed 2–3 to a cage and fasted overnight at 23 °C (6 p.m. to 10 a.m.). Overnight-fasted mice were administered an oral glucose load of 2 g/kg body weight (41.8–47.8 mg for WT mice, 76.8–94.4 mg for ob/ob mice). To measure blood glucose and insulin levels, 15 μL blood was collected from the tail veins at 0, 2, 5, 10, 15, 20, 30, 45, 60, 90, 120, 180, and 240 min after glucose administration (n = 5). We used the blood glucose and insulin levels measured in our previous study^{12 (link)} (Fig. S2). For the metabolome and transcriptome studies, mice were sacrificed at 0, 20, 60, 120, and 240 min after glucose administration, and the gastrocnemius muscle was excised. Muscle samples were frozen immediately in liquid nitrogen and homogenized with dry ice. The powdered samples were divided and used for metabolomics, lipidomics, transcriptomics, a glycogen assay, and western blotting.