Experiments were carried out in Transwell 24-insert plate chambers (Corning, NY, USA). The cell migration experiment was performed by seeding 5×10 4 shTMEM16A and ANO1 LV cells in 100 µL of medium with 1% FBS (Gibco-Invitrogen, Carlsbad, CA, USA) into the upper compartment of the chamber with a polycarbonate lter (6.5-mm diameter, 8-mm pores; Corning Costar, Corning, NY, USA). The lower chambers contained 600uL of medium containing 10% FBS (Gibco) to serve as chemoattractant. For the cell invasion experiment, Transwells were coated-with Matrigel (BD Biosciences, Franklin Lakes, NJ, USA). The migrated or invaded cells in the lower surface of the lters were xed with methanol, stained with Giemsa (Coolaber, Beijing, China) and counted under a light microscope (Olympus Corp, Tokyo, Japan) in 5 randomly selected visual elds at magni cation of 400×.
Transwell 24 insert plate chambers
Manufactured by Corning
Sourced in United States
The Transwell 24-insert plate chambers are a laboratory equipment designed for cell culture and migration studies. The product consists of a 24-well plate with individual permeable membrane inserts that allow for the separation of two cell culture compartments. This setup enables researchers to study various cellular processes, such as cell migration, invasion, and co-culture experiments, in a controlled and reproducible manner.
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2 protocols using transwell 24 insert plate chambers
1
Cell Migration and Invasion Assay
2
Cell Invasion Assay Protocol
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Cell invasion activity was evaluated in 8.0 μm pore size transwell 24-insert plate chambers (Corning, Acton, MA, USA) coated with BioCoatMatrigel (BD Biosciences, Bedford, MA, USA). After pre-starved for 24 hours by culturing in serum free medium, 3X104 (NCI-H520) or 5X104 (GLC82) cells per well were plated in the upper chambers with serum free medium and incubated for 48 (NCI-H520) or 72 (GLC82) hours. The lower chambers were filled with 10% fetal bovine serum medium. After wiping the cells off from the upper side of the upper chamber, the lower side of the upper chamber was fixed with methanol and stained with 4,6-diamidino-2-phenylindole (DAPI; Sigma). Images were photographed under microscope. The DAPI staining area in the lower chamber was normalized to scrambled shRNA group or DMSO group, indicating its relative invasiveness.
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