Figure 3. CAV1-phosphorylation on tyrosine 14 during cell spreading on pure ECM surfaces.

In spreading assays, B16F10(CAV1/wt) cells (1,5×10⁶) were allowed to attach to A. fibronectin, B. laminin, C. vitronectin and D. collagen IV-coated plates (2 μg/ml) for different periods of time (0, 5, 15, 30 and 60 min), with time 0 representing cells in suspension. Then, whole cell lysates were prepared and pY14-CAV1 levels were determined by Western blotting. Upper graphs show the densitometric analysis of relative pY14-CAV1 levels during cell spreading. Lower images show pY14-CAV1, CAV1 and Actin (control) expression by Western Blotting. Lower panels show cells in phase contrast and stained with phalloidin during spreading. The average area per cell is indicated in μm². Data shown are the averages from three independent experiments (mean ± S.E.M, n=3,***p<0.001; **p<0.01 and *p<0.05).