Trpa1 ko mice

All experiments were performed according to the ethical guidelines recommended by the Kyoto University Animal Research Committee. The protocol was approved by the Kyoto University Animal Research Committee (permit number: 2014–45, 2015–38). All efforts were made to minimise the number of animals used and limit experimentation to only what was necessary. Male ICR mice aged 6–7 weeks were purchased from Japan SLC (Shizuoka, Japan). TRPA1-KO mice were obtained from Jackson Laboratory (Bar Harbor, ME, USA) and genotyped as previously described23 (link). TRPV1-KO mice were a gift from Dr. David Julius (University of California, San Francisco, USA). Each KO mouse line was backcrossed with C57BL/6J mice for 7 to 10 generations to eliminate any background effects on the phenotypes. All mice were housed under constant ambient temperature (24 ± 1 °C) and humidity (55 ± 10%), with alternate light–dark cycles from 8:00 a.m. to 8:00 p.m. Food and water were freely available.

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)).