Fig. 5: Adult Ripk1cko/K376R mice are hypersensitive to TNF-induced death.
From: Impaired RIPK1 ubiquitination sensitizes mice to TNF toxicity and inflammatory cell death
a Dosing scheme for deleting the WT allele in Ripk1K376R/cko Rosa26-Cre.ERT2 or Ripk1wt/cko Rosa26-Cre.ERT2 animals by tamoxifen [80 mg/kg, IP dosing]. b Histology of liver, spleen, and small intestines after 28 days of dosing to induce recombination [scale bar 100 μm liver, 50 μm small intestine, 200 μm spleen]. c Kaplan–Meier survival curve of mice after TNF [500 μg/kg iv injections] induced SIRS model at d11 after induced deletion. Difference between the two groups by Mantel–Cox test: P < 0.0001. d Ripk1cko/+ and Ripk1cko/K376R BMDMs, treated in vitro with 4-OH-tamoxifen [30 nM] for 5 days, were treated with TNF [100 ng/ml], zVAD [20 μM] and GNE684 [5 μM]. RIPK1-IP was performed and analyzed by western blot for complex II formation. e Percentage Sytox Green positive BMDMs, indicating membrane permeabilization and cell death, detected over 8 h by Incucyte with the indicated treatments [TNF – T [5 ng/ml], BV6 – B [500 nM], zVAD – Z [20 μM] and GNE684 [5 μM]. The mean of three independent experiments with SD is plotted. Asterisks indicate statistical analysis between treated Ripk1cko/+ and Ripk1cko/K376R samples. P values, repeated measures two-way ANOVA followed by Tukey’s multiple comparison test (*** > 0.0002).
