Figure 2

Adhesion of cultured VSMC to Netrin-4. (A) Crystal violet staining of cells that adhered to the substrate. 96-well plates were coated overnight with BSA, Vitronectin (VN; 2.5 μg/ml) or Netrin-4 (N4, 1 μg/ml). VSMC were added to the wells and incubated for 4 hours at 37°C. BSA was used as a negative control and Vitronectin (VN) was used as a positive control. VSMC adhere to Netrin-4 (N4) or Vitronectin but not to BSA. (B) Quantification of cell adhesion with the three substrates by measurement of absorbance. All assays were performed in triplicate in at least three different experiments. (C) Contrast phase image of VSMC adhesion in the presence of Vitronectin, 20x magnification (D) Contrast phase image of VSMC adhesion in the presence of Netrin-4, 20x magnification. Scale bar: 50 μm.