SNU449 and Huh7 in which ZNF545 was unexpressed and re-expressed were suspended in serum-free medium. Cells (2×105) were placed into the upper chamber of an 8-um pore size transwell apparatus (Corning, NY, USA) and incubated for 30 hours. Cells that migrated to lower surface of the membrane were stained with crystal violet and counted in three independent high-power fields (×200). For invasion analysis, SNU449 and Huh7 in which ZNF545 was unexpressed and re-expressed (2×105) were seeded into the upper chamber of a transwell apparatus coated with extracellular matrix (ECM) gel (BD Biosciences, San Jose, CA) and incubated for 48 hours. Cells that invaded the lower membrane surface were stained with crystal violet and counted in three independent high-power fields (×200). The migration and invasion assay of HXBF344 cells (1×105) were performed as mentioned above, and the incubated time were 10 hours and 48 hours respectively.
Extracellular matrix ecm gel
Manufactured by BD
Extracellular matrix (ECM) gel is a complex mixture of structural and functional proteins that mimic the natural extracellular environment. It provides a three-dimensional, biologically active scaffold for cell culture and tissue engineering applications.
Automatically generated - may contain errors
2 protocols using extracellular matrix ecm gel
1
Transwell Migration and Invasion Assay
2
Cell Cycle, Migration, and Invasion Assays
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KYSE150 and TE1 cells in which NKD2 was unexpressed and reexpressed were starved 12 hours for synchronization, and the cells were restimulated with 10% fetal bovine serum for 24 hours. Cells were fixed with 70% ethanol and treated using the Cell Cycle Detection Kit (KeyGEN Biotech). The cells were then sorted by a FACS-Caliber flow cytometer (BD Biosciences, Mansfield, MA). The cell phase distribution was analyzed by Modfit software (Verity Software House, Topsham, ME).
Transwell Assay KYSE150 and TE1 cells in which NKD2 was unexpressed and reexpressed were suspended in serum-free medium. Cells (2 Â 10 5 ) were placed into the upper chamber of an 8-mm pore size transwell apparatus (Corning, Corning, NY) and incubated for 20 hours. Cells that migrated to the lower surface of the membrane were stained with crystal violet and counted in three independent high-power fields (Â200). For invasion analysis, KYSE150 and TE1 cells in which NKD2 was unexpressed and reexpressed (2 Â 10 5 ) were seeded into the upper chamber of a transwell apparatus coated with extracellular matrix (ECM) gel (BD Biosciences, San Jose, CA) and incubated for 36 hours. Cells that invaded the lower membrane surface were stained with crystal violet and counted in three independent high-power fields (Â200).
Transwell Assay KYSE150 and TE1 cells in which NKD2 was unexpressed and reexpressed were suspended in serum-free medium. Cells (2 Â 10 5 ) were placed into the upper chamber of an 8-mm pore size transwell apparatus (Corning, Corning, NY) and incubated for 20 hours. Cells that migrated to the lower surface of the membrane were stained with crystal violet and counted in three independent high-power fields (Â200). For invasion analysis, KYSE150 and TE1 cells in which NKD2 was unexpressed and reexpressed (2 Â 10 5 ) were seeded into the upper chamber of a transwell apparatus coated with extracellular matrix (ECM) gel (BD Biosciences, San Jose, CA) and incubated for 36 hours. Cells that invaded the lower membrane surface were stained with crystal violet and counted in three independent high-power fields (Â200).
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