Migration of HCmel12-eGFP cells was quantified using transwell assays. Briefly, 5 × 104 cells in 250 μl of RPMI 1640 with 1% FCS were placed in the upper chamber of uncoated polyethylene terephthalate (PET) filters (8 μm pore size, BD Biosciences) in a 24-well plate and subsequently incubated at 37°C/5% CO2 to adhere. Six hours later, FR was added at the concentrations indicated in Fig. 1. RPMI containing 1% FCS was added to the lower chamber. Twenty-four hours later, cells in the upper chamber were removed by a cotton swab. Transmigrated cells on the lower surface of the membrane were counted in three high-power fields (magnification, ×100) using a TE Eclipse microscope (Nikon).

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