Fig. 2: K612R mutation in mRIPK1 disrupts the formation of complex I and complex II downstream of TNFR1 activation by TNFα. | Nature Communications

Fig. 2: K612R mutation in mRIPK1 disrupts the formation of complex I and complex II downstream of TNFR1 activation by TNFα.

From: Ubiquitination of RIPK1 regulates its activation mediated by TNFR1 and TLRs signaling in distinct manners

Fig. 2

a, b Complex IIa was immunoprecipitated by FADD antibodies and analyzed by western blotting in Ripk1+/+ and Ripk1K612R/K612R MEFs pretreated with SM-164 (100 nM) (a) or 5Z-7 (200 nM) (b) for 2 h, and then treated with mTNFα (100 ng/ml) for different time points in the presence or absence of Nec-1s (10 μM) as indicated. Uncropped blots in the Source Data file. c, d Complex IIb was immunoprecipitated by RIPK3 antibodies and analyzed by western blotting in Ripk1+/+ and Ripk1K612R/K612R MEFs pretreated with SM-164 (100 nM) (c), 5Z-7 (200 nM) (d) for 2 h, and then treated with mTNFα (100 ng/ml)/Z-VAD (25 μM) for indicated time. Uncropped blots in the Source Data file. e Complex I in Ripk1+/+ and Ripk1K612R/K612R MEFs treated with mTNFα (100 ng/ml) for indicated time points was immunoprecipitated with TNFR1 antibody and analyzed by western blotting. Uncropped blots in the Source Data file. f, g Complex I in Ripk1+/+ and Ripk1K612R/K612R MEFs pretreated with SM-164 (100 nM) (f) or 5Z-7 (200 nM) (g) for 2 h, and then treated with flag-mTNFα (100 ng/ml) for indicated time points was immunoprecipitated with Flag M2 Agarose Affinity gel and analyzed by western blotting in. Uncropped blots in the Source Data file.

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