24-well Transwell chambers with an 8 mm pore size (Corning Incorporated, NY, USA) were precoated with Matrigel (BD Biosciences, Bedford, MA). 600 μL medium supplemented with 10% FBS was added to each lower chamber. 3×104 cells in 200 μL serum-free medium were seeded in each upper chamber and cultured for 24 h. Fixed with 4% paraformaldehyde for 15 min and stained with 0.1% crystal violet for 15 min, cells invaded to the lower surface of the membrane were pictured and counted in 5 random fields (200× magnification).

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