FIG. 1.

The effect of MC160 expression on TNF-α-induced expression of an NF-κB transcriptionally regulated luciferase gene. (A and B) 293T cells were cotransfected with 450 ng of pNF-κBluc, 50 ng of pRL-null, and 500 ng of either pCI or MC160/pCI. (A) After 24 h, cells were incubated in medium lacking or containing TNF (0 to 20 ng/ml) for 4 h and then harvested. Their lysates were analyzed for luciferase activity. (B) Cells were incubated with medium lacking or containing 10 ng TNF/ml, and cells were harvested either 4, 6, 8, or 12 h later. Their lysates were then analyzed for luciferase activity. (C) Cells were transfected with reporter plasmids and the indicated amounts of either pCI or a combination of pCI and MC160/pCI. Following a 24-h incubation, cells were incubated in medium lacking or containing TNF (10 ng/ml) and were harvested 4 h later. Lysates were analyzed for luciferase activities. An asterisk (*) indicates statistically significant inhibition (P < 0.05). For all experiments, 10 μg of each cellular lysate was screened for MC160 expression by immunoblotting (IB) with anti-MC160 protein antisera, as illustrated by the immunoblot inset above each graph.