Figure 4.

Endothelial cells plated on osteopontin have elevated NF-κB binding activity. (A) Cells were plated on polylysine (PDL) or osteopontin (OPN)-coated surfaces in serum-free medium. 4 h after plating, nuclear protein extracts were harvested, and EMSA was performed using a double-stranded ³²P-labeled consensus NF-κB oligomer. Adding a 100-fold excess of unlabeled consensus NF-κB oligomer (COMP) completely inhibited binding, and a 100-fold excess of unlabeled unrelated sequence (UNCOMP) had no effect. (B) Cells were plated on osteopontin (OPN), fibronectin (FN), and collagen type I (COLL I). 4 h after plating, nuclear protein extracts were harvested, and EMSA was performed using a double-stranded ³²P-labeled consensus NF-κB oligomer. (C) Cells were plated on polylysine (PDL) or osteopontin (OPN)-coated surfaces in serum-free medium. Nuclear protein extracts were harvested at 2, 4, 6, and 8 h after plating, and EMSA was performed using a double-stranded ³²P-labeled consensus NF-κB oligomer.