ADK deficiency results in spontaneous RIPK1-driven liver damage. (A–D) Adk ^f/f, Adk^f/f;Alb-Cre, or Adk^f/f;Alb-Cre;Ripk1^D138N/D138N mice were analyzed. In the unstressed conditions at the age of 7 wk, the levels of p-RIPK1(S166), CC8, CC3, and ADK in hepatocytes isolated from the livers were determined by immunoblotting (A); liver H&E staining, TUNEL staining, and p-RIPK1(S166) immunostaining were performed (B); serum ALT/AST detection (C) was performed. Scale bar: 100 μm. The survival time of mice of different genotypes was analyzed (D). (E–H) Vehicle, 5-ITu (100 mg/kg), and Nec-1s (20 mg/kg) were intraperitoneally injected to 8-wk-old Ripk1^WT/WT or Ripk1^D138N/D138N 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). Scale bar: 100 μm. The survival time of mice of different treatments was analyzed (H). Data are represented as the mean ± SD (B, C, F, and G). Data are representative of n = 3 independent experiments (A–H). Statistical significance was determined using one-way ANOVA with post hoc Dunnett’s test (B, C, F, and G) or two-sided log-rank test (D and H). ***P < 0.001. Source data are available for this figure: SourceData F7.