The necroptotic pathway plays a primary role in TNF-induced death of Ripk1 ^S161E/S161E mice. (A and B) Survival curves and body temperature of 8- to 12-wk-old male littermate mice of the indicated genotypes after TNF injection (50 μg/kg, i.v.). (C and D) Representative photos and H&E staining images of ceca from three mice of each indicated genotype (50 μg/kg TNF treated for 6 h, i.v.). Scale bars, 100 μm. (E) Mice of the indicated genotypes were i.v. injected with TNF (50 μg/kg). At the indicated time points, serum samples were collected for measurements of LDH. Data are presented as mean ± SD. P values are determined by a two-tailed, unpaired t test. ***P < 0.001. All mice were 8- to 12-wk-old male littermates (C–E). (F and G) Survival curves of 8- to 12-wk-old male littermate mice of the indicated genotypes after i.v. injection with TNF of the indicated doses. Mouse survival is presented as a Kaplan–Meier plot, and the log-rank test is performed. ns, P ≥ 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Data of body temperature are presented as mean ± SD. The above experiments were independently performed twice.