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

Figure 6. IL-1 signaling in different TAB- and TRAF6-deficient cells.

Figure 6.

(A) Generation of TAB1/TRAF6 DKO cells from TAB1 KO cells (clone 44 from Figure 5A). (B) WT TRAF6 and the E3 ligase-inactive TRAF6[L74H] and TRAF6[120-522] mutants were re-expressed in TAB1/TRAF6 DKO cells (clone 1 from A). These cells, TAB1 KO cells (clone 44 from Figure 5A) and TAB1/TRAF6 DKO cells not re-transfected with TRAF6, were then stimulated with IL-1β for the times indicated. Other details are as in A. (C) HA-TAB2 or the K63-Ub-binding-defective HA-TAB2[T674/F675A] mutant were re-expressed in TAB1/2/3 TKO IL-1R* cells (clone A4 from Figure 1A). These cells, TAB1/2/3 TKO cells not re-transfected with HA-TAB2 and WT IL-1R* cells, were stimulated for 10 min with 5 ng/ml IL-1β. Cell extracts (20 µg of protein-WT cells or 40 µg of protein-TAB1/2/3 TKO cells) were subjected to SDS–PAGE and immunoblotted with the antibodies indicated.