Fig. 5.

ZBP1 homointeraction is important in triggering RIPK3-dependent necroptosis. a, b WT L929 or RIPK3 KO L929 cells were infected with lentiviral vector to express proteins as indicated, and cell survival was determined at 36 h after infection by measuring the ATP levels. c A total of 293T cells were transfected with plasmids as indicated. TCL and anti-FLAG immunoprecipitates were immunoblotted to detect the indicated proteins. d A total of 293T cells were transfected with plasmids as indicated. TCL and anti-FLAG immunoprecipitates were treated with or without β-mercaptoethanol (β-Me) and immunoblotted to detect the indicated proteins. e WT L929 cells were infected with lentiviral vectors to express proteins as indicated and were then treated with PBS (ctrl), ethanol (ETOH), or 4-OHT for 12 h; cell survival was then determined by measuring the ATP levels. f WT L929 cells were infected with lentiviral vectors to express proteins as indicated. The FLAG and GAPDH protein levels were measured by western blotting. g A total of 293T cells were transfected with plasmids as indicated, and sequential immunoprecipitation with anti-Flag antibody and anti-HA antibody was performed. TCL and the anti-FLAG and sequential anti-HA immunoprecipitates were immunoblotted to detect the indicated proteins. For a, b, and e, the data are from n = 3 independent experiments. The error bars indicate the mean ± s.e.m. of n = 3 independent experiments. A two-tailed Student’s t-test was applied to determine the P values. **P < 0.01. For c, d, f, and g, the results shown are representative of the results of three independent experiments