Cell migration was detected using a scratch-wound assay 20. Briefly, the 90% confluent cell layer was scratched using a 10-µl pipette tip, and detached cells were removed by washing with PBS. The cells were cultured in medium with 2% FBS for 48 h. Images of the scratch wounds were obtained using an inverted phase-contrast microscope (BX53, Olympus Corporation, Tokyo, Japan) equipped with a digital camera at 0, 24 and 48 h. The wound width was determined using Image-Pro Plus software version 6.0.

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