To investigate the effects of PSH on cell migration, a wound healing assay was performed. Cells were seeded into 6-well plates at 1×106 cells/ml, and cultured until a confluent monolayer was formed. The cell surface was wounded by a 200-µl pipette tip. PBS was used to remove the cells from the wound area, and the cells were treated with different doses of PSH in DMEM containing 1% FBS; the serum concentration was based on previous studies (23,24). Images of the wound areas were captured by an inverted microscope (×200 magnification) at the start of the experiment (0 h) and following treatment for 72 h. For each experiment, five replicates were performed, and the experiment was repeated three times. The gap width was calculated using ImageJ software (version 1.42), and the width was expressed in arbitrary units. The wound healing rate was calculated as follows: Wound healing (%)=(1-B/A) × 100%, where A is the gap width at 0 h, and B is the gap width at 72 h.

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