B6.129x1 trpv1tm1jul j

Adult male TRPV1^−/− (B6.129X1-Trpv1^tm1Jul/J) mice and their genetically unaltered wild type (WT) counterparts were purchased from Jackson Laboratory, Maine, USA, and bred in the animal house located in the School of Pharmacy as per protocols approved by the IACUC and as per the ethical regulations. Mice were housed in a climate-controlled environment (22.8 ± 2.0 °C, 45–50% humidity) with a 12/12-light/dark cycle with access to designated diet and water ad libitum. Weight gain, food and water intake were monitored weekly. Starting from week 6, mice were housed in groups of four in separate cages and randomly assigned into feeding groups of NCD or HFD (± CAP) until week 38. At the end of 38 weeks, metabolic study was performed and brown fat pads were obtained and used for in vitro experiments.
Subgroups of WT mice were fed NCD (± CAP) for 32 weeks. We performed a dose response for three concentrations of CAP (0.003%, 0.01% and 0.03% w/w in HFD) to determine their effect on weight gain. For experiments performed with mice to determine the effect of CAP supplementation post HFD, we fed WT and TRPV1^−/− mice HFD for 16 weeks (week 6 through 22) followed by supplementation of CAP in HFD (0.01% of CAP) from week 22 through 38. The weight gain and food/water intake were monitored weekly and metabolic studies were performed on weeks 16 and 32.

All animal procedures were in accordance with guidelines by the National Institutes of Health, and approved by the Institutional Animal Care and Use Committee at Columbia University. Wild-type, CB2-deficient (B6.129P2-Cnr2tm1Dgen/J, Jax stock number 005786), and Trpv1-deficient mice (B6.129X1-Trpv1tm1Jul/J, Jax # 003770) were purchased from Jackson Laboratories. FAAH mice were a gift from Benjamin F. Cravatt, Scripps Research Institute, La Jolla.^{17 (link)} CB1-deficient mice were generated by breeding CB1^fl/fl mice (a gift from Beat Lutz, University of Mainz, Germany) with germ line-deleter Ella-Cre (B6.FVB-Tg (EIIa-cre) C5379Lmgd/J, Jax # 003724). All knockout mice were intercrossed with C57Bl/6 mice, followed by establishment of knockout mice and wild-type controls from heterozygote mice.

TRPV1^Cre/Cre (Trpv1tm1(cre)Bbm, 017769, Jackson Laboratory) were mated with ROSA26-tdTomato^fl/fl (B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J, 007909, Jackson Laboratory) to yield TRPV1^Cre/+/ROSA26-tdTomato^fl/+ mice. Female TRPV1^−/− mice were mated with male TRPV1^−/− mice (B6.129X1-Trpv1tm1Jul/J, 003770, Jackson Laboratory). Female TRPA1^−/− mice were mated with male TRPA1^+/− mice (B6;129P-Trpa1tm1Kykw/J, 006401, Jackson Laboratory). Genotype of the offspring was confirmed using polymerase chain reaction. Wild type C57BL/6J mice were purchased from Jackson Laboratory (000664). All experiments were performed with approval from the University of South Florida Institutional Animal Care and Use Committee (AAALAC #000434).

Female C57BL/6 mice and TLR4-mutant C₃H/HeJ (Tlr4^Lps-d) mice, 8–12 weeks old were purchased from National Cancer Institute (Frederick, MD). Female vanilloid receptor knockout mice on BL/6 background (B6.129X1-Trpv1^tm1Jul/J), and mast cell-deficient mice (WBB6F1/J-Kit^W/Kit^W-v) and their WT (+/+) littermate controls were purchased from The Jackson Laboratory (Bar Harbor, ME). Mice were housed under standard pathogen-free conditions in the Animal Resource Facility of University of South Carolina School of Medicine and all experiments were conducted after obtaining prior approval from the Institutional Animal Care and Use Committee.