3.2.5. Migration and Invasion Assay

For migration assay, a scratch assay was performed. Each melanoma cells (2.0 × 10⁵ cells per well) were seeded in 24-well plates. After 24 h, cells were scratched with a SPLScar^TM Scratcher (SPL Life Sciences, Pocheon, Korea) and the detached cells were removed by PBS washing twice. Cells were incubated in complete media with 0.01 μM concentrations of each compound for 12 h. The images were acquired at 0 h, and 12 h incubation with 100× magnification, and percent of migration were accessed using ImageJ (n = 3).

For invasion assay, Boyden chamber assay was conducted using QCM ECMatrix Cell Invasion Assay kit (ECM 550, Sigma-Aldrich, Munich, Germany) according to the manufacturer’s instruction. Briefly, cells were seeded in the transwell chamber insert (8 μm pore size) at a density of 5.0 × 10⁵ cells per well after serum starvation for 12 h. The cells were incubated with 0.01 μM concentration of each compound for 24 h at 37 °C. The non-invaded cells were eliminated and followed the invaded cells staining. Cells were observed with 100 × magnification. Stained cells were dissolved in 10% acetic acid. After transferring the 96-well plate, optical density at 560 nm was measured by EnVision^® 2105 microplate reader (PerkinElmer, Waltham, MA, USA) and quantified % of invaded cells (n = 3).