To evaluate the ability of cell migration, the transwell assays were preformed using 24-well transwell chambers with 8 µm pore membranes (Corning LifeSciences). Incubated in serum-free medium for 24 h, 20,000 cells were seeded into in the top chamber of the insert, by followed adding media containing 10% FBS in the lower chamber as a chemoattractant. The chambers were cultured at 37 °C for 48 h, washed twice with PBS, and fixed in 100% methanol on the ice for 15 min. Then the cells were stained with 0.05% crystal violet for 15 mins, followed by carefully cleaning away the cells in the top chamber of the insert. Chose 5 fields randomly and count for analyzing. All experiments were performed in triplicate.
To evaluate the ability of cell invasion, the transwell assays were preformed using 24-well transwell chambers with 8 µm pore membranes (Corning LifeSciences). The transwell chambers for invasion assays must be pre-coated with Matrigel (BD Biosciences, San Jose, CA, USA). The next steps are the same as above.
To evaluate the ability of cell invasion, the transwell assays were preformed using 24-well transwell chambers with 8 µm pore membranes (Corning LifeSciences). The transwell chambers for invasion assays must be pre-coated with Matrigel (BD Biosciences, San Jose, CA, USA). The next steps are the same as above.