NF-𝜿B and Matrix-Dependent Regulation of Osteopontin Promoter Activity in Allylamine-Activated Vascular Smooth Muscle Cells

Figure 3

Expression of Rel proteins in control and allylamine cells seeded on plastic, collagen I, fibronectin, and laminin. Protein extracts from cells at 80% confluence were electrophoresed in a 7% denaturing polyacrylamide gel, transferred onto nitrocellulose, and blocked overnight in 5% milk in TBS with 0.1% Tween. Membranes were probed overnight followed by incubation with horseradish peroxidase-labeled secondary antibody for 4 hours and then incubated with 1 : 2 chemiluminescent substrate: TTBS for 5 minutes, followed by visualization through autoradiography. Similar results were seen in three separate experiments. C: control; A: allylamine.