4.4. Wound Healing Assay

B16F10 cells were seeded in a six-well plate and allowed to attach overnight to 80% confluency. Subsequently, cell monolayers were wounded by pipette tips and washed with PBS twice to remove floating cells. Cells were treated with or without different concentrations of RT for up to 48 h. Cells migrated into the wound surface and the number of migrating cells was determined under an inverted microscopy at various times; five randomly chosen fields were analyzed for each well. The percentage of inhibition was expressed using untreated wells at 100%. Three independent experiments were performed.