FIGURE 3.

Melanoma in the presence of PFP reduced VE-dependent HUVEC adhesion in a B-Raf(V600E)- and thrombin-dependent manner. A, HUVEC monolayers were left alone or co-cultured with WM35, A375M, UACC903, or Lu1205 cells in the presence or absence of PFP. After being detached, HUVECs were plated onto a VEC-Fc- or Fc-coated dish in the presence of respective melanoma-HUVEC co-culture medium for 60 min. Cell adhesion was measured as described under “Experimental Procedures” and is shown as the percentage of adherent cells. Values are mean ± S.E. from three independent experiments. B and C, after co-culture with Lu1205 and UACC903 cells in the presence or absence of PFP, HUVECs were plated onto a VEC-Fc-coated dish for the time indicated. Cell adhesion is shown as the percentage of adherent cells. D, HUVECs were co-cultured with Lu1205 cells transfected with buffer, scrambled siRNA, siB-Raf(V600E), or siTF, or treated with vehicle or hirudin in the presence of PFP. Then they were assessed for adhesion activity as described in the legend to A. E, HUVECs transfected with buffer, scrambled siRNA, or siPAR-1 were co-cultured with Lu1205 cells in the presence of PFP. Then they were assessed for adhesion activity as described in the legend to A. *, p < 0.05. N.S., not significant.