CD151 affects endothelial cell spreading, chemotaxis, invasion, and random migration. (A) To assess spreading, MLECs were seeded onto plates coated with fibronectin, gelatin, or Matrigel. After 30 minutes, spread cells were counted in 3 high-power fields in duplicate wells (*P < .01). Data are representative of 3 independent experiments with similar results. (B) For chemotactic migration, MLECs (7 × 104 cells in 200 μL DMEM; 5% FCS) were plated in upper Transwell chambers, coated with fibronectin, gelatin, or Matrigel (thin layer). Bottom chambers contained 600 μL DMEM, 5% FCS, and 20 ng/mL bFGF. Cells migrating through the filter were counted (*P < .05; **P < .01). (C) To assess invasion, cells were counted after migrating for 6 hours through a Matrigel-coated Transwell (n = 3; *P < .005). (Di) To measure random migration, cells were plated on coverslips coated with Matrigel or gelatin. (ii) Cell movements were recorded by time-lapse video microscopy every minute for 30 minutes and quantified using Scion Image tools (n = 12). Top panels show randomly selected individual migration tracks copied and combined (bars = 100 μm). Bottom panels show “T” and “D/T” where T is total distance migrated and D is distance between starting and ending point (*P < .05). High D/T ratios (approaching 1.0) indicate directional persistence.32 Data are representative of 3 independent experiments with similar results.
