Ripk1 ^R606K/R606K mice exhibited mitigated RIPK1-driven liver IRI and SIRS. (A–G) Ripk1 ^WT/WT or Ripk1^R606K/R606K mice underwent 1-h ischemia/6-h reperfusion operation. The levels of RIPK1 R606me2s, p-RIPK1(S166), CC8, CC3 in isolated hepatocytes were determined by immunoblotting (A). Serum ALT/AST detection (B) and liver H&E staining (C) were performed; liver TUNEL staining (D), CC3 immunostaining (E), MPO immunostaining (F), and serum proinflammatory cytokine detection (G) were performed. Scale bar: 100 μm (C–F). (H–J) 500 μg/kg TNFα was intravenously injected to Ripk1^WT/WT or Ripk1^R606K/R606K mice. Body temperature (H) and survival time (I) were compared. NF-κB and MAPK activation in liver was analyzed with immunostaining (J). Data are represented as the mean ± SD (B–H). Data are representative of n = 3 independent experiments (A–J). Statistical significance was determined using two-way ANOVA with post hoc Bonferroni’s test (B–H) or log-rank test (I). ***P < 0.001. Source data are available for this figure: SourceData F10.