FIGURE 6.

Mal interaction with TRAF6 is required for TLR2- and TLR4-mediated transactivation of the p65 subunit of NF-κB. Immortalized Mal-deficient macrophages were virally reconstituted with either GFP-tagged Mal (lanes 2 and 7–9) or MalE190A (lanes 3 and 10–12) as described previously in Fig. 5. GFP-positive populations were challenged with either 10 ng/ml of Pam3Cys (A) or LPS (B) for indicated time points and lysed, and protein was separated by SDS-PAGE followed by immunoblot (IB) for p65, p65 Ser²⁷⁶, or p65 Ser⁵³⁶ phosphorylation where indicated. The results represent three independent experiments.