For the migration assay, 50 000 cells per well (triplicate) were seeded in medium without serum onto 8 μm polypropylene filter inserts in Boyden chambers (Fisher). Medium with 10% FBS was added onto the well. After 24 h, cells on the bottom of the filter were fixed and stained with Thermo Scientific Shandon Kwik Diff Stains (Fisher). The invasion assay was done as described above, except that the 8 μm polypropylene filter inserts were coated with Matrigel (#CB‐40234; Corning Inc., Corning, NY, USA) and incubated for 24 h. Ten visual fields were randomly chosen under microscopy and cells were quantified by using imagej software (National Institutes of Health, Bethesda, MD, USA).
Thermo scientific shandon kwik diff stains
Manufactured by Thermo Fisher Scientific
The Thermo Scientific Shandon Kwik Diff Stains are a set of reagents used for the rapid staining of blood films and other cytological preparations. The stains provide a quick and consistent means of differentiating cell types for microscopic examination.
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Cell Migration and Invasion Assay
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Cell Migration and Invasion Assay
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For the migration assay, 50,000 cells per well (triplicate) were seeded in medium without serum onto 8-μm polypropylene filter inserts in Boyden chambers (Fisher). Medium with 10% FBS was added onto the well. After 24 h, cells on the bottom of the filter were fixed and stained with Thermo Scientific Shandon Kwik Diff Stains (Fisher). The invasion assay was done as described above, except that the 8-μm polypropylene filter inserts were coated with Matrigel (#CB-40234, Corning, USA) and incubated for 24 h. Ten visual fields were randomly chosen under microscopy and cells were quantified by using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
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