FIG. 9.

Dominant negative NIK inhibits pIC-stimulated NF-κB-dependent transcription T98G cells were transiently cotransfected with an NF-κB-dependent luciferase reporter (5 NF-κB sites) and 500 ng of a plasmid expressing a kinase-dead mutant of NIK or the corresponding empty vector as the control. Approximately 36 h posttransfection, cells were treated with pIC or TNF-α (100 μg/ml and 10 ng/ml, respectively, for 8 h) and extracts were prepared and analyzed for luciferase activity. The graph reflects firefly luciferase units corrected for transfection efficiency by expression from a Renilla luciferase plasmid included as an internal control in the dual-luciferase assay system. The experiment was performed in duplicate and is representative of several separate experiments.