Fig. 5.

Functional ablation of endogenous TRAF6 by ODC/AZ system. HEK293T cells were transiently transfected with a reporter gene plasmid (0.1 μg) that contains a NFκB-responsive element cloned upstream of a luciferase reporter gene, with 0.01 μg of pCMVβ-gal as a transfection-efficiency control and 0.1 μg of the indicated plasmids encoding ODC, ODC-TRAF6C, ODC-RANKp, or AZ, in various combinations as indicated (total DNA amount normalized). After 24 h, cells were treated with 50 ng/ml IL-1 (Left) or 10 ng/ml TNFα (Right) for an additional 24 h, and luciferase activity was measured in cell lysates and normalized relative to β-galactosidase (mean ± SD; n = 3).