Extended Data Fig. 3: Ripk1D325A/D325A Mlkl−/− Fadd−/− mice develop splenomegaly and lymphadenopathy similar to Mlkl−/− Fadd−/− mice.
From: Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis
a, Western blots of 293T cells overexpressing mouse caspase-8 and mouse RIPK1 (left), primary MEFs (middle), or BMDMs (right). Results representative of two independent experiments. The N-terminal RIPK1 cleavage product appears larger in the 293T cells owing to the N-terminal Flag tag. T, TNF; C, cycloheximide. β-Actin loading control performed after MYC (left) or caspase-8 (middle, right). For gel source data, see Supplementary Fig. 1. b, E12.5 littermates representative of six Ripk1D325A/+ Ripk3−/− and four Ripk1D325A/D325A Ripk3−/− embryos. c, E10.5 embryo sections immunolabelled for either RIPK1 or caspase-8 (brown). Results representative of five wild-type and six Casp8−/− embryos. Scale bar, 100 μm. Asterisk shows maternally derived Casp8+/− decidua in the placenta. d, Schematic showing the organization of the Fadd knockout allele. e, Leukocyte numbers in mice aged 10–12 weeks (wild-type, n = 2; Mlkl−/−, n = 3; Mlkl−/− Fadd−/−, n = 4; Ripk1D325A/D325A Mlkl−/− Fadd−/−, n = 4). Circles, different mice. Lines, mean ± s.e.m. f, Flow cytometric analysis of mesenteric lymph node cells from the mice in e. Mean ± s.e.m.
