Imaging of cell morphological changes was performed 3 days after treatment with FR, trametinib, or vehicle by bright-field microscopy. Cells were grown at a density of 15,000 cells per well for HCmel12, 92.1, and Mel285 cells or at a density of 30,000 cells per well for Mel270, Mel290, Mel202, and OMM1.3 cells in RPMI medium on 96-well ibidi plates coated with poly-d-lysine (PDL) for high-resolution microscopy. Compounds or vehicle was added 3 hours after seeding, and the plate was incubated for another 72 hours at 37°C/5% CO2. Cells were then fixed with 4% paraformaldehyde (PFA) for 20 min at room temperature and washed twice with phosphate-buffered saline (PBS). Images were taken with a Leica DFC 360FX microscope.

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