Melanoma cells (5 × 104) were seeded in duplicate in serum-free DMEM into an 8-μm pore Transwell membrane (Corning) coated with Matrigel (BD Biosciences). Invasion analysis was performed as previously described (13). Images of fixed cells from the invasion chambers were taken using an Olympus IX81 microscope and the cellSens Dimension software. In all assays, three fields per insert were photographed. The number of invaded cells was normalized to the number of total seeded cells.

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