FIG. 7.

MKK73E-induced activation of the IL-8 promoter is suppressed by coexpression of dominant negative NIK. HEK-293 cells were transfected with 5 μg of pCS3MT-MKK73E, pCS3MT-MKK73A, pCS3MT-NIK, or pCS3MT-NIK(KK429-430AA) in the indicated combinations together with pUHC13-3-IL-8pr. Where required, empty pCS3MT was added to keep total DNA amounts constant. (A) At 48 h after transfection, cells were lysed and luciferase reporter gene activity was determined as for Fig. 6 (mean ± SEM from three independent experiments); (B) 100 μg of lysate proteins from one experiment was separated by SDS-PAGE and Western blotted to confirm equal expression of Myc-epitope tagged protein kinases MKK7 and NIK. RLU, relative light units.