Extended Data Fig. 2: Characterization of mice expressing c-FLIP(D371A/D377A) or CYLD(D215A).
From: Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis
a, Western blots of thymocytes that were freshly isolated (−), or cultured in either medium alone or FasL for 2.5 h. β-Actin loading control performed after caspase-8. Results representative of two independent experiments. b, Kaplan–Meier survival curve of Cflar+/+ (n = 10) and Cflar2×DA/2×DA littermates (n = 14). P = 0.3, two-sided log-rank test. c, Graph shows the percentage of thymocytes that were viable 24 h after treatment. Circles, cells from different mice (Cflar+/+, n = 3; Cflar2×DA/2×DA, n = 3). Bars, mean ± s.e.m. Unpaired two-tailed t-test. d, Flow cytometric analysis of mesenteric lymph node cells of wild-type (n = 2) and Cflar2×DA/2×DA (n = 2) littermates, and as controls, Fadd−/− Mlkl−/− mice (n = 2). Numbers below each genotype indicate total lymph node cellularity. Analyses used the gating strategy shown in Extended Data Fig. 11. e, Schematic showing the organization of the CyldD215A allele. f, Western blots of thymocytes. Arrowheads highlight caspase-dependent CYLD cleavage products. β-Actin loading control performed after cleaved caspase-8. Results representative of two independent experiments. g, Littermate males aged 18 days. Results representative of 29 wild-type and 23 CyldD215A/D215A mice. h, Kaplan–Meier survival curve of Cyld+/+ (n = 29) and CyldD215A/D215A (n = 23) littermates. P = 0.9, two-sided log-rank test. For gel source data, see Supplementary Fig. 1.
