Pre-treatment of fibronectin and SMC-derived ECM with NE decreases VEGF binding. A: 96-well hydrophobic plates were coated with 10 μg/ml fibronectin in the presence of 10 μg/ml heparin. Plates were treated with NE then 125I-VEGF (0.45 nM) was added for 2 h. Bound VEGF was released with 2 M NaCl, 25 mM HEPES, pH 7.5. B: Primary rat smooth muscle cells were grown 5 days past confluence, cell layer dissolved, and the exposed extracellular matrix was incubated with NE. ECM coated wells were then incubated with 125I-VEGF (0.45 nM) and bound VEGF extracted. Each data point is the mean of quadruplicate determinations ± SEM. Regression analyses with ANOVA of both dose response curves revealed a statistically significant effect of NE treatment with Significance F and p values below 0.05.
Forsten-Williams et al. BMC Systems Biology 2011 5:170 doi:10.1186/1752-0509-5-170