Fig. 3: Ripk1K376R/K376R reduces TNF-induced NF-κB and MAPK signaling and promotes cell death complex formation.
From: Impaired RIPK1 ubiquitination sensitizes mice to TNF toxicity and inflammatory cell death
a Western blot and b FLAG-TNF immunoprecipitation (IP) of the same experiment of Ripk1+/+ and Ripk1K376R/K376R primary MEFs after treatment with 1 μg/ml FLAG-TNF for the indicates times, examined with indicated antibodies. c Western Blot of primary MEFs and IP after treatment with TNF [100 ng/ml], zVAD [20 μM] and GNE684 [5 μM] for the indicated time points. d Percentage of dead primary MEFs [Sytox Green positive cells] compared to positive control. Signal was measured every hour for 20 h in the Incucyte with the indicated treatments: TNF [T] 100 ng/ml, zVAD [Z] 20 μM, BV6 [B] 500 nM, GNE684 5 μM. The Mean value with SD for six independent experiments is shown. Asterisks indicate statistical analysis between treated WT and K376R samples. After 9 h for TNF and TB treatment and after 5 h for TZ, the significance did not change. P values, repeated measures two-way ANOVA followed by Tukey’s multiple comparison test (* > 0.0332, ** > 0.0021, *** > 0.0002).
