Investigating NLRP3 Inflammasome in Mice

All mice used in this study were male mice on C57BL/6 background at ages indicated in specific experiments. Nlrp3^R258W (+/R258W) mice and Nlrp3^−/− mice were reported previously^{21 (link), 53 (link)}. Nlrp3^R258W mice were maintained as heterozygotes by backcrossing with WT (+/+) control mice from the Jackson Laboratory. All Nlrp3^R258W and WT mice used in this study are littermate controls. Il1r1^−/−, Rag1^−/−, and Casp1/11^−/− mice were from the Jackson Laboratory. Nlrp3^eGFP reporter mouse line was reported before^{24 (link)}. All mice were maintained in specific pathogen-free facility under a strict 12 h light cycle (on at 0700 hours and off at 1900 hours), and given a regular chow diet ad libitum (#M01-F25, Shanghai SLAC Laboratory Animal Co.). Cohousing (ch-) of WT and Nlrp3^R258W mice were carried out at 1:1 ratio since weaning (3–4 weeks) till adulthood (10–12 weeks), or the mice were separated upon weaning according to genotype, denoted as singly housed (sh-). Germ-free mice on C57BL/6 background were purchased from Shanghai SLAC Laboratory Animal Co., and maintained in a germ-free facility. All animal experiments were performed in compliance with the guidelines from the national care and use of laboratory animals and were approved by the institutional ethics committee of the Institut Pasteur of Shanghai, Chinese Academy of Sciences.