A Transwell chamber was placed in a 24-well plate, and 800 μl of ECM medium containing 10% FBS was added to each well. A total of 2×104 cells were diluted in 200 μl of serum-free ECM medium and were added to the upper chamber. Following incubation at 37°C for 24 h, the upper surface of the membrane was wiped with a cotton-tipped applicator to remove non-migratory cells and the migratory cells on the under surface were fixed with 10% formaldehyde and stained with 2% crystal violet (Beijing Solarbio Science & Technology Co., Ltd.) solution for 10 min at room temperature. The membranes were mounted on glass slides and the number of migrated cells from 10 random microscopic fields were counted by manual counting after obtaining images. The image data were collected using a Canon EOS 600d camera (Canon EOS 600d).

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