For analysis of cell proliferation, HCmel12 cells (1.5 × 105 per well) and UM cell lines (2 × 105 per well) were seeded into six-well plates in complete RPMI medium. Six hours later, FR or MEK inhibitor trametinib (1 μM) was added. Reference groups were solvent-treated. Cell growth was photo-documented over time using a Zeiss Axio Vert.A1 inverted microscope. After 72 hours, cells were collected and viable cells were counted using trypan blue.

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