A549-stable clones over-expressing TCF-4K or pEGFP-N1 empty vector as a control (4 x 10 5 ) were suspended in 200 µL serum-free medium and seeded into the upper chamber of a transwell insert with an 8 µm pore size membrane (Corning Costar Corp., Cambridge, MA, USA). RPMI 1640 containing 10% FBS was placed in the lower chamber as a chemoattractant. After incubation for 12 h, the migrated cells on the underside of the filter membrane were fixed and stained with 0.1% crystal violet. The number of migrated cells was counted in 8 randomly selected microscopic fields and photographed. The protocol used for the invasion assay was the same as that used for the migration assay, except that the transwell insert was coated with Matrigel (BD Biosciences, Heidelberg, Germany).
Transwell insert with an 8 m pore size membrane
Manufactured by Corning
Sourced in United States
The Transwell insert with an 8 µm pore size membrane is a laboratory equipment product. It is a cell culture insert designed to allow the passage of cells, molecules, or other materials through a semipermeable membrane with a pore size of 8 micrometers.
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2 protocols using transwell insert with an 8 m pore size membrane
1
Transwell Assay for Cell Migration and Invasion
2
Transwell Migration and Invasion Assay
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Stable SATB1-overexpressing, SATB1-knockdown, and empty vector control cells (1×105) were suspended in 200 µL serum-free medium and seeded into the upper chamber of a transwell insert with an 8-µm pore size membrane (Corning Costar Corp, Cambridge, MA, USA). DMEM containing 10% FBS was placed in the lower chamber as a chemoattractant. After incubation for 24 h, non-migrated cells in the upper chamber of the transwell insert were removed with a cotton swab, and the migrated cells on the underside of the filter membrane were fixed and stained with 0.1% crystal violet. The number of migrated cells was counted in 5 randomly selected microscopic fields and photographed. The protocol used for the invasion assay was the same as that used for the migration assay, except that the transwell insert was coated with Matrigel (BD Biosciences, Heidelberg, Germany).
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