Casp1 11 mice

C57BL/6, Nlrp3^-/-, and Casp1/11^-/- mice were obtained from Jackson Laboratory (Bar Harbor, ME). Asc^-/-, TLR11^-/- and Myd88^-/- mice have been previously described [16 (link), 52 (link), 53 (link)]. TLR11^-/- mice were crossed with Casp1/11^-/- mice to generate TLR11xCasp1/11^-/-. All control and experimental mice were age- and sex-matched within all individual experiments. This study included both male and female mice, and the data derived from male and female mice identified no sex-specific differences in the performed experiments. All mice were maintained at in the pathogen-free American Association of Laboratory Animal Care-accredited animal facility at the University of Rochester Medical Center, Rochester, NY. All animal experimentation was conducted in accordance with the guidelines of the University of Rochester’s University Committee on Animal Resources (UCAR), the Institutional Animal Care and Use Committee (IACUC).

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.