Fig. 3.

Cells bearing RIPK1^K376R/K376R are more susceptible to TNF-α induced apoptosis and necroptosis. a Ripk1^+/+ and Ripk1^K376R/K376R MEFs were treated with T, TS, TSZ, TC, TCZ for indicated times, respectively. Cell viability was determined using the CellTiter-Glo kit. The data are represented as the mean ± SEM of MEFs derived from 3 embryos. P values were determined by Student’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001). Abbreviations are as follows: UT untreated, TSZ TNF-α (20 ng/ml)+Smac mimetic(100 nM) +zVAD(20 μM); TSZN, TNF-α+Smac mimetic +zVAD+Necrostatin-1 (20 μM); TC, TNF-α+CHX (20 ng/ml); TCZ, TNF-α+CHX +zVAD. Source data are provided as a Source Data file. b Ripk1^+/+ and Ripk1^K376R/K376R MEFs were treated with mouse TNF-α (20 ng/ml) for indicated times, the cell lysates were analyzed by western blotting using the indicated antibodies. c Ripk1^+/+ and Ripk1^K376R/K376R MEFs were treated with TSZ or TSZN for 3 h, the cell lysates were resolved on non-reducing gel and immunoblotted with anti-p-MLKL antibody. d Ripk1^+/+ and Ripk1^K376R/K376R MEFs were treated with mouse TNF-α (20 ng/ml) for 12 h, anti-RIPK1 was used to immunoprecipitate complex and immunocomplexes were analyzed by western blotting using indicated antibody