RIPK1 R606K mutation protects mice against ADK inhibition–induced RIPK1-driven liver injury. (A and B) Illustration of the generation of Ripk1^R606K/R606K mice (A) and the genotype as determined by Sanger sequencing (B) are shown. (C) Number of offspring from intercrossing of Ripk1^WT/R606K mice. (D) Body weight of male and female mice of Ripk1^WT/WT or Ripk1^R606K/R606K genotypes at the age of 2 mo. (E–K) Vehicle or 5-ITu (100 mg/kg) was intraperitoneally injected to Ripk1^WT/WT or Ripk1^R606K/R606K mice every day. 72 h after the first injection, the levels of p-RIPK1(S166), CC8, CC3, and ADK in hepatocytes isolated from the livers were determined by immunoblotting (E); serum ALT/AST detection was performed (F); liver H&E staining, TUNEL staining, and p-RIPK1(S166) immunostaining were performed (G); liver CC3 and p-RIPK3 immunostaining (H) and CD11b immunostaining (I) were conducted; mRNA levels of Tnf, Il6, Il1β, Ccl2 (J) were analyzed. The survival time of mice of different groups was analyzed (K). Scale bar: 100 μm (G–I). Data are represented as the mean ± SD (D and F–J). Data are representative of n = 3 independent experiments (A–K). Statistical significance was determined using two-tailed unpaired Student’s t test (D), one-way ANOVA with post hoc Dunnett’s test (F–I), two-way ANOVA with post hoc Bonferroni’s test (J), or log-rank test (K). **P < 0.01; ***P < 0.001. Source data are available for this figure: SourceData F9.