Transwell membranes 24 well

Transwell membranes (24-well; Costar, Cambridge, MA, USA) were coated with Matrigel for 6 h for the invasion assay or without Matrigel for the migration assay. In total, 2 × 10⁵ cells in serum-free medium were seeded onto the upper chamber, and 750 μL of medium with 10% FBS was added to the lower chamber. After incubation for 24 h (for migration) and 48 h (for invasion), the cells adhering to the upper surface of the membrane were removed with a cotton swab. The invasion or migration cells, which adhered to the lower surface, were stained with crystal violet and counted in 4 representative fields by light microscopy (×40 magnification).

Transwell membranes (24-well, Costar, Cambridge, MA, USA) were either coated with Matrigel for 6 h for invasion assays or were used without Matrigel for migration assays. Cells (0.5 × 10⁵) in serum-free medium were seeded into each upper chamber, and 600 μl medium supplemented with 10% FBS were added to each lower chamber. After incubation for 24 h, cells adhering to the upper surface of the membrane were removed with a cotton swab. Cells that had invaded or migrated, which were adhered to the lower surface, were stained with 0.1% crystal violet and counted in four representative fields by light microscopy (200 × magnification).

Transwell membranes (24-well; Costar, Cambridge, MA, USA) were coated with Matrigel for 6 h for the invasion assay or without Matrigel for the migration assay. In total, 2 × 10⁵ cells in serum-free medium were seeded onto the upper chamber, and 750 μL of medium with 10% FBS were added to the lower chamber. After incubation for 24 h (for migration) and 48 h (for invasion), the cells adhering to the upper surface of the membrane were removed with a cotton swab. The invasion or migration cells that adhered to the lower surface were stained with crystal violet and counted in four representative fields using light microscopy (×40 magnification).