Culture medium was added to the lower wells of Boyden chambers (BD Bioscience), and cells were added to the upper chambers. Migratory cells attached to the lower side of the inserts were fixed with 4% PFA, permeabilized with 0.2% Triton X-100, and stained with Phalloidin (Life Technologies). The inserts were washed with 1× PBS (Life Technologies) and imaged using a 4× objective on an inverted Olympus IX 71 epifluorescence microscope and analyzed with ImageJ. Matrigel invasion assays were performed using Matrigel-coated 24-well transwell plates (BD Biosciences) as described above.
Matrigel coated 24 well transwell plates
Manufactured by BD
Matrigel-coated 24-well transwell plates are a type of cell culture insert designed for use in migration and invasion assays. The plates feature a semi-permeable membrane coated with a basement membrane extract, providing a 3D environment for cell studies.
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2 protocols using matrigel coated 24 well transwell plates
1
Cell Migration and Invasion Assay
2
Matrigel Invasion Assay Protocol
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Matrigel invasion assays were performed using Matrigel-coated 24-well Transwell plates (BD Biosciences). DMEM complete media (10% FBS) was added to the lower chambers, and 1 × 10 5 A549E and A549M cells in DMEM with 0.5% FBS were added to the upper chambers. After 24 h, non-migratory cells on the top surface of the filters were wiped off with cotton balls. The cells on the top surface of the filters were wiped off with cotton balls, and migratory cells attached to the lower side of the inserts were fixed with 4% paraformaldehyde (PFA) and was stained with DAPI. Finally, migrated cells were counted under a fluorescence microscope at 10× magnitude (Axiovert 200; Carl Zeiss, Jena, Germany).
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