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. 2000 Apr 11;97(9):4615–4620. doi: 10.1073/pnas.080583397

Figure 1.

Figure 1

Dominant-negative IKK-2 blocks PMA- and Raf-mediated NF-κB activation. (A) Jurkat T cells were cotransfected with a κB-dependent reporter (3xκB.luc) and either an expression vector for dn-IKK-2 or the empty expression vector (pcDNA3). Sixteen hours after transfection, cells were stimulated with PMA (20 ng/μl) or TNF-α (80 ng/μl) for 6 h as indicated. The luciferase activity for the noninduced cells cotransfected with pcDNA3 was arbitrarily set to 1, all other activities are given relative to this standard. (B) Jurkat T cells were cotransfected with 3xκB.luc and expression vectors for dn-IKK-2, Raf-BXB-CX, and Raf-BXB-DD as indicated. Stimulation with PMA was performed as above. (C) Jurkat cotransfection with the HSV-tk.luc reporter and expression vectors for either wild-type IKK-2 or dn-IKK-2. Expression level of the HSV-tk.luc construct (10 μg) cotransfected with the empty expression vector (pcDNA3) was set to 100%. (D) Jurkat cells were transfected with the κB-dependent reporter and active Raf as indicated. Sixteen hours post transfection, the cells were treated with suramin (1.5 mM f.c.). One hour later, PMA or TNF-α was added where indicated, and cells were harvested 6 h later.