NF-κB activation by plasmin.
(A) Kinetic study. Monocytes were stimulated with plasmin 0.43 CTA U/mL or LPS 1 μg/mL (positive control) for the times indicated. Cells were lysed, and nuclei were isolated. Nuclear extracts (5 μg) were subjected to EMSA with a 32P-labeled DNA probe containing the NF-κB binding site. (B) Supershift and competition study. Monocytes were stimulated with plasmin 0.43 CTA U/mL for 1 hour. Nuclear extracts were incubated for 1 hour with anti-p65, anti-c-Rel, anti-p50, and anti-p52 antibodies or with unlabeled NF-κB or AP-2 specific oligonucleotides (100-fold molar excess). Supershift assay revealed binding of p65, c-Rel, and p50, but not of p52 nuclear factor. Preincubation with NF-κB–specific oligonucleotide sequence abolished formation of the NF-κB complex. AP-2–specific oligonucleotides had no effect on the NF-κB/DNA complex formation. Results of 1 of 3 experiments are shown.
