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. 2017 Jun 26;474(13):2235–2248. doi: 10.1042/BCJ20170288

Figure 3. TRAF6 E3 ligase activity is required for the IL-1β-dependent activation of the TAB1–TAK1 heterodimer in TAB2/3 DKO cells.

Figure 3.

(A) TAB2/3 DKO (clone 4 from Figure 1A) and TAB2/TAB3/TRAF6 TKO IL-1R* cells were stimulated with IL-1β for the times indicated, and cell extracts subjected to SDS–PAGE and immunoblotting with the antibodies used in Figure 1 and an antibody recognizing IRAK4 phosphorylated at Thr245 and Ser346. (B) As in A, except that WT TRAF6 or the indicated E3-ligase-inactive mutants of TRAF6 were re-expressed in the TAB2/TAB3/TRAF6 TKO IL-1R* cells. (C) WT and TAB2/3 DKO cells were incubated for 72 h with siRNA against Ubc13 and stimulated for 10 min with 5 ng/ml IL-1β. Other details are as in A.