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. 2016 Jan 22;113(6):1612–1617. doi: 10.1073/pnas.1518163113

Fig. S4.

Fig. S4.

The DUB CYLD can associate with DCT-NEMO in response to TNF. (A) 293 cells were transfected with CYLD, full-length NEMO, or E391X-NEMO vectors and treated with TNF for 10 min before immunoprecipitation of cell lysates with anti-NEMO followed by Western blot for CYLD and NEMO in IP and whole-cell lysates. (B) Reconstituted Jurkats were stimulated with TNF for 10 min, lysed, and CYLD was immunoprecipitated with anti-CYLD. The upper band in the NEMO blot of the IP is a combination of mouse heavy chain and full-length NEMO. High molecular-weight RIP1 is stabilized at the TNFR in cells that express E391X-NEMO. (C) Quantitation by densitometry of Western blot bands corresponding to RIP1 (unmodified and ubiquitinated) coimmunoprecipitated with TNFR1 as performed in Fig. 4D, (n = 3). The effect of genotype (NEMO or E391X) was determined by two-way ANOVA and was highly significant (P < 0.0001).