Effect of AT III on NF-κB activation in monocytes and ECs.

Cells were either untreated or pretreated for 2 hours with the indicated concentrations of AT III, followed by a 1-hour stimulation with either LPS (monocytes; 10 μg/mL) (A) or TNF-α (ECs; 40 ng/mL) (B) in the presence and absence of AT III. Nuclear proteins from monocytes and ECs were analyzed by EMSA for binding to an oligonucleotide containing the NF-κB consensus sequence. Representative photographs of EMSA gels are shown in the upper panels. In the lower panels, a quantitation of the inhibiting effect of AT III on the agonist-induced binding activity is given. Band intensities corresponding to the NF-κB–DNA complexes were quantified by densitometric analysis using a Bio-Imaging analyzer. Values represent means ± SEM from 9 independent experiments (each cell type). *P < .01 versus stimulation in the absence of AT III.