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. Author manuscript; available in PMC: 2012 Jun 1.
Published in final edited form as: Nat Immunol. 2011 Nov 6;12(12):1176–1183. doi: 10.1038/ni.2157

Figure 8.

Figure 8

Reconstitution of Cyld−/− BMDMs with WT but not Cyld(Y485A) mutant rescues defects in termination of Tak1 activation and chronic production of inflammatory cytokines. (a) V5-Cyld(WT) or V5-Cyld(Y485A) mutant was ectopically expressed in Cyld−/− BMDMs using a lentiviral transduction system. The cells lysed at indicated time points post TNF stimulation. The lysate was immunoprecipitated using antibody against TRAF2 and immunoblotted with antibody against p-Tak1. The same membrane was reprobed with antibody against Tak1. The cell lysate from WT BMDMs stimulated with TNF was used as control in these experiments. (b) WT and Cyld−/− transduced with V5- Cyld(WT) or V5-Cyld(Y485A) mutant were stimulated with TNF and expression of IL-6, TNF and IL-1β was analyzed by real-time PCR. (c) IL-6 and IL-1β concentration were assayed in the culture supernatant by ELISA. Data are representative of at least three independent experiments (b–c mean and s.d. of triplicate wells).