Figure 3.

Inhibition of NF-κB function in AdvIκBα-infected macrophages. M-CSF-treated monocytes were infected with AdvO or AdvIκBα at a moi of 50. A third group was untreated. After an additional 3 days in M-CSF the cells were washed, and half of each group was treated (+) or otherwise (−) with 10 ng/ml LPS for 30 min. (A) The cells then were lysed, and cytosolic and nuclear extracts were prepared and analyzed for IκBα/β expression in the cytosol, or p65/p50 expression in the nucleus, by immunowestern blotting. Protein (150 μg and 50 μg) was analyzed for each cytosol or nuclear sample, respectively. (B) Nuclear extracts were prepared and 20 μg protein analyzed for NF-κB (Upper) or AP-1 (Lower) activity by electrophoretic mobility-shift assay.