Fig. 4

CD39 abrogates ATP induced nuclear factor (NF)-κB translocation.

Confluent human umbilical vein endothelial cells (HUVEC) were left untreated or infected with CD39 or β-galactosidase adenovirus, and subsequently, stimulated with ATP or ATPγS. Nuclear proteins were extracted from HUVEC and equal amounts of nuclear extracts were incubated with a radiolabeled NF-κB-specific oligonucleotide. Resulting DNA/protein complexes were separated by electrophoretic mobility shift assay (EMSA) on a 6% polyacrylamide gel. Controls with competing unlabeled or mutated probes for NF-κB confirmed specificity. One experiment, representative of three, is shown and confirms that CD39 up-regulation blocked the ATP-mediated NF-κB translocation. NI, non-infected; rAdCD39, recombinant replication-deficient adenovirus from human CD39 cDNA; rAdβgal, recombinant replication-deficient adenovirus from bacterial β-galactosidase.