HPAF-II (1.5 × 105 /well) and CL-901 (105/well) cells were suspended in serum-free culture in the upper compartment of transwell chambers (8.0 μm pore polycarbonate membrane insert-Corning Inc., NY, USA) pre-coated with 30 μg/well of Matrigel Reduced Growth Factors (Corning). Fusion proteins (1 μM) were also added in the upper compartment of the transwell. The lower compartment of the chamber for HPAF-II cells was filled with 2% FBS and HGF (12.5 ng/ml), while for CL-901 was added cell culture medium supplemented with 1% FBS and HGF (25 ng/ml). After 24 h, cells on the upper side of the transwell filters were mechanically removed, while cells migrated through the membrane were fixed with 11% glutaraldehyde and stained with 0.1% crystal violet. All images were captured with optical microscopes (Leica, Wetzlar, Germany). Images were quantified with Image-J software.

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