Effects of platelets on hepatocellular carcinoma cell motility and invasion. (A) Huh7-GFP cells were assayed by Matrigel chemo-invasion with BSA 1% (control) or different dilution of platelet factors added to a lower compartment of the chamber at the indicated concentrations. (B) Tracking the migration of Huh7-GFP cells by live optical imaging. GFP-expressing Huh7 were imaged for 16 h and the behavior of motile cells was recorded. (C) The tracks of 6 representative cells (3 controls and 3 stimulated with platelet factors) are plotted inWind-rose plots with the initial position of each track (left panel).Scale bar, 100 μm. The temporal increase in the productive motility of these representative tracks from controls and stimulated Huh7-GFP cells are also plotted in the graph (right panel). (D) 3D Invasion assay of Huh7-GFP cells in the presence of platelet soluble factors. The rate of cell migration was determined at different time intervals (6, 12, 24 h). Three representative tracks for each condition (cells stimulated with platelet factors or with BSA 1% as control) are plotted in the graph. All experiments were carried out in triplicate. The results are expressed as mean ± SEM. P values were determined by Mann–Whitney one-tailed test.