Tnfr1ko

All procedures were executed in compliance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and according to the University of Miami Institutional Animal Care and Use Committee (IACUC) approved protocol (Protocol #: 21-070). TNFR1 and Gsdmd knockout animals (TNFR1KO and GsdmdKO, respectively; these knockouts have the C57BL/6 J genetic background) and C57BL/6J mice as the wild-type (WT) controls were received from the Jackson Laboratory (Bar Harbor, ME, USA; stock numbers 003242, 032663, and 000664). The oral iron chelator deferiprone (DFP, 1 mg/mL, #379409, MilliporeSigma, St. Louis, MO, USA) was delivered to animals via drinking water. We used 2–4-month-old male and female mice to address sex as a biological variable. Animals were housed under standard conditions of humidity and temperature, were given free access to food and water, and had a 12-h light to dark cycle. All methods were completed and reported in accordance with ARRIVE guidelines.

All procedures were performed in compliance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and according to the University of Miami Institutional Animal Care and Use Committee (IACUC) approved protocols. Tnfr1, Ifnar1, and Gsdmd knockout animals (Tnfr1KO, Ifnar1KO, and GsdmdKO, respectively) and C57BL/6 J mice as the wild type controls were obtained from the Jackson Laboratory (Bar Harbor, Maine, United States; stock numbers 003242, 028288, 032663, 000664). Mice were housed under standard conditions of temperature and humidity, with a 12-h light to dark cycle and free access to food and water. All animals have the C57BL/6J genetic background. All methods were completed and reported in accordance with ARRIVE guidelines.

Rik1^K376R/K376R mutation mice were generated by CRISPR-Cas9 technique. Briefly, one single guide RNA (sgRNA) that target the DNA region surrounding K376 of RIPK1 and the corresponding donor construct (containing K to R mutation) were designed (see in Supplementary Table 1). To prevent the sgRNA-directed Cas9 from cleaving mutated RIPK1 genomic locus, we also constructed several synonymous mutations within the donor. Then, the donor constructs, the mRNA of the sgRNA and Cas9, were injected together into the fertilized eggs of C57BL/6 mice. KD-Ripk1^−/− mice was also generated by CRISPR-Cas9 technique following the similar process above. The corrected founder mice were genotyped by genomic PCR and DNA sequencing (see in Supplementary Table 2). WT C57BL/6 mice, TNFR1 KO, RIPK3, and Caspase8-KO mice were purchased from Jackson Laboratory and bred in the facility. All mice were housed in the specific pathogen-free animal facilities in Tsinghua University. All mouse experiments were performed in compliance with institutional guidelines and according to the protocol approved by the Institutional Animal Care and Use Committee of Tsinghua University.