FIGURE 4.

VWF multimer strings recruit breast cancer cells to EC surfaces under shear stress. HUVEC monolayers were cultured on Ibidi channel slides and stimulated with PMA to allow maximum release of VWF. MDA‐MB‐231, fluorescently labeled with Cell Tracker green were flown over the HUVEC layer at flow rate of 0.2 ml/min resulting in venous shear stress of 0.25 dyn/cm². (A) MDA‐MB‐231 cells binds to VWF strings under shear flow. (B) The specificity of this interaction was confirmed using a blocking anti‐VWF antibody. (C) LMWH significantly decreased the number of binding events observed. (D) Equilibrium surface coverage (cells per mm²) and (E) detachment rates (K ^off) were calculated for each treatment condition. Perfusion assays were performed on HUVEC monolayers stimulated with supernatant from MDA‐MB‐231 cells or supernatant from MDA‐MB‐231+platelets and adhesion of labeled MDA‐MB‐231 cells were quantified (F and G). Data are presented as the mean ± SEM of at least three independent experiments. Data were analyzed with analysis of variance (anova) followed by Bonferroni t ‐test and p values < .05 were considered to be significant (*p < .05, **p < .005, ***p < .001). Scale bar 100 μm.