FIGURE 5.

Disruption of TRAF-6-dependent NEMO-IKK-2 interactions in cell-free system by NBD peptides. A, HeLa cell extracts were prepared and treated with 0, 0.25, 0.5, 1, or 2 mm ATP with or without 60 nm exogenous TRAF6. B and C, HeLa cell extracts were treated with 0.1 or 1% DMSO (vehicle), PHA-408, WT, or mutant NBD peptides (0.1 or 1% DMSO, final concentration) in the absence or presence of 2 mm ATP and 60 nm exogenous TRAF6 and were analyzed by Western blot. The experiments were repeated three times with similar results. HSP27 was used as a control for protein loading. NBD peptides used were IKK-2 11-mer.