Fig. 6

RI-3 prevents in vitro adhesion to endothelium and trans-endothelial migration of melanoma cells. a HUVEC were seeded onto matrigel and allowed to attach and grow for 24 h (CTRL) prior to seeding GFP-A375 cells suspended in complete endothelial medium plus diluents (None), or 10 nM RI-3 at 37 °C, 5% CO₂. At the indicated times, cell associated fluorescence was assessed by a fluorescence plate reader. Data represent means ± SD of three independent experiments performed in duplicate. Statistical significance with ***p < .0.0001. b After 2 h, cells were stained with rhodamine-phalloidin and GFP-A375 cells (arrows) visualized on multiple z-series collected at 0.20 μm intervals by laser confocal microscopy. On the left representative images recorded in 3D projection are shown. Original magnifications: 400×. c Trans-endothelial migration of A375 cells. HUVECs (1 × 10⁴ cells/well) suspended in growth medium, were grown until they formed a confluent monolayer, prior to seeding A375 cells (1 × 10⁴ cells/well) in growth medium plus diluents (None) or 10 nM RI-3. Data represent mean ± SD from a quadruplicate experiment