Genetic Manipulation of IRAK2 in Obesity

IRAK2-deficient and IRAK2 kinase-inactive gene-targeted mice were previously described^{23 (link), 24 (link)}. Myd88^fl/fl (008888), IL-1R1-deficient (003245), Adiponectin-Cre (028020), and Ucp1-luc2, Ucp-tdTomato (026690)^{50 (link)} mice were purchased from The Jackson Laboratory. HA-tagged IRAK2 reporter mice were generated by inserting 3xHA expressing cascade upstream the stop codon of Irak2 gene (Cyagen Biosciences). HA-tagged MyD88 reporter mice were generated by inserting 3xHA expressing cascade upstream the stop codon of Myd88 gene (Cyagen Biosciences). IRAK2 conditionally deficient mice by flanking exon 1 of Irak2 by loxP sites (Cyagen Biosciences). 8-week old male mice were maintained on a high-fat diet composed of 60% kcal derived from fat (TD 06414, Envigo) for 12 weeks, while non high-fat diet mice maintained on either standard rodent chow (2918 Teklad Global 18% Protein Rodent Diet, Envigo). 6-weeks old mice were used to isolate adipose tissues for primary adipocyte culture. Animals were housed in a specific pathogen-free facility with a 14-h light/10-h dark cycle and given free access to food and water. All procedures using animals were approved by the Cleveland Clinic Institutional Animal Care and Use Committee (Protocol Number: 2018–1814).