|Incubate 24 hours in a tissue culture incubator.
Cell invasion assays were performed with the 24-well cell invasion assay kit (Sigma-Aldrich) according to the manufacturer’s instructions. Cells (n=900) were resuspended in culture medium without FBS and placed in the upper Transwell chamber in triplicate. After a 24-h incubation at 37°C (5% CO atmosphere), the cells in the upper surface of the membrane were removed using a cotton swab and the cells that had migrated to the lower surface of the membrane were fixed and stained. The migrated cells on the lower surface of the membrane filter were scored from six random fields under a CKX31 microscope (magnification, ×400; Olympus, Tokyo, Japan). The results are shown as percentage migrating rate (%): % = (migrated cells/total cells) × 100. The wound healing assay was performed according to the manufacturer’s instructions. To generate a wound field, cells were cultured until they formed a monolayer around the insert. Subsequent to removing the insert, an open wound field was generated and cells were able to migrate from either side of the gap. Full paper
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