The upper chamber of the Transwell chamber was covered with Matrigel (50 mL; 356,234; Becton, Dickinson and Company, NJ, USA). The transfected cells were then seeded into the upper cavity of the Transwell filter membrane and incubated for 48 h in serum-free medium. The infiltrated cells on the underside of the filter membrane were fixed with 5% glutaraldehyde, stained with 0.1% crystal violet and counted under the microscope. The invasion was determined by the number of cells that passed through the Matrigel.

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