8 m pore size chambers

To analyze wound healing, SKOV3 cells were seeded in 6-well plates at a density of 5×10⁵ cells/well with RPMI. When the cells reached 80% confluency, they were infected with the ILK or NC lentivirus. After 48 h, the cell monolayer was wounded using a plastic pipette tip. Cells were subsequently rinsed with PBS and cultured with serum-free RPMI for 24 h at 37°C. The wound closure was observed and imaged under a phase-contrast microscope (Olympus Corporation, Tokyo, Japan). For the Transwell assays, 8-µm pore size chambers (Corning, Inc., Corning, NY, USA) were used with or without an insert coated with Matrigel (BD Biosciences, San Jose, CA, USA). A total of 48 h after infection with the ILK or NC lentivirus at 37°C, 1×10⁵ cells in serum-free medium were added to the upper chamber. The lower chamber was filled with 10% FBS RPMI. After 24 h of incubation at 37°C, the cells remaining on the upper surface of the membrane were removed, whereas the cells that had invaded through the membrane were fixed with 100% methanol for 15 min at room temperature and stained with 0.1% crystal violet for 20 min at room temperature. Images of SKOV3 cells were obtained under a phase-contrast microscope (Olympus Corporation).

For the Transwell assays, 8-µm pore size chambers (Corning, Inc.) were used with an insert without (migration) or with (invasion) Matrigel coating (BD Biosciences). At 24 h after transfection, 1x10⁵ cells in serum-free medium were added to the upper chamber. The lower chamber was filled with 10% FBS RPMI 1640. After 24-h incubation, cells remaining on the upper surface of the membrane were removed, whereas cells that had invaded through the membrane were fixed with 0.1% paraformaldehyde for 20 min at room temperature, stained with 0.1% crystal violet for 30 min at room temperature, imaged and counted under a light microscope (magnification, x10).