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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 6.

Figure 6

Inhibiting Tak1 activation rescues elevated inflammatory cytokine production by Itch−/− and Cyld−/− BMDMs (a) BMDMs were pre-incubated with indicated amounts of (5Z)-7-Oxozeaenol(Ox) and stimulated with TNF for 6 hours, expression of IL-6, TNF, and IL-1β was analyzed by real-time PCR. (b) BMDMs were pretreated with 100 nM of Ox for 30 minutes and then stimulated with TNF for the indicated time points. The cell lysates were immunoprecipitated using antibody against TRAF2 and immunoblotted using antibody against p-Tak1. The total lysate was immunoblotted using antibodies against TRAF2 and actin. (c) WT, Itch−/− and Cyld−/− BMDMs were transduced with a lentiviral vector encoding dominant negative Tak1 (dnTak1). The efficiency of ectopic expression was analyzed by immunoblotting the total cell lysate using antibody against V5. (d) WT, Itch−/− and Cyld−/− BMDMs transduced with dnTak1 were stimulated with TNF and expression of IL-6, TNF and IL-1β was analyzed by real-time PCR. Data are representative of three experiments. (a, d mean and s.d. of triplicate wells).