Cell migration assay was performed in 24-well transwell plate with 8-mm polyethylene terephalate membrane filters (Corning) separating the lower and upper culture chambers. In brief, H1299 or A549 cells were plated in the upper chamber at 5 × 104 cells per well in serum-free DMEM medium. The bottom chamber contained DMEM medium with 10% FBS. Cells were allowed to migrate for 24 h in a humidified chamber at 37 °C with 5% CO2. After the incubation period, the filter was removed and non-migrant cells on the upper side of the filter were detached using a cotton swab. Filters were fixed with 4% formaldehyde for 15 min and cells located in the lower filter were stained with 0.1% crystal violet for 20 min and photographed.
8 mm polyethylene terephalate membrane filters
Manufactured by Corning
The 8-mm polyethylene terephalate membrane filters are a type of lab equipment designed for filtration purposes. These filters are made of polyethylene terephalate, a durable and reliable material. The core function of these filters is to separate and remove particles or contaminants from liquids or gases passing through them.
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3 protocols using 8 mm polyethylene terephalate membrane filters
1
Cell Migration Assay in Transwells
2
Transwell Assay for Cell Migration
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Cell migration assays were performed in 24-well transwell plates with 8-mm polyethylene terephalate membrane filters (Corning) separating the lower and upper culture chambers. In brief, LN229 or U251 cells were plated in the upper chamber at 1 × 104 cells per well in serum-free DMEM medium. The bottom chamber contained DMEM medium with 10% FBS. Cells were allowed to migrate for 24 h in a humidified chamber at 37 °C with 5% CO2. After the incubation period, the filter was removed and non-migrant cells on the upper side of the filter were detached using a cotton swab. Filters were fixed with 4% formaldehyde for 15 min and cells located in the lower filter were stained with 0.1% crystal violet for 20 min and photographed.
3
Transwell Migration Assay for Smurf1 Depletion
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5 × 104 control or Smurf1-depleted HCT116 cells were added in the upper well of the 24-well transwell plate with 8-mm polyethylene terephalate membrane filters (Corning) in 200 μL serum-free RPMI 1640 medium supplemented with 1 μM GEM or 6.25 μM CIS. The bottom reservoir was filled with 500 μL RPMI 1640 medium plus with 10% fetal bovine serum (FBS) and 100 U mL-1 penicillin. Cells were cultured at 37°C in a humidified 5% CO2 incubator. After 24 h, cells on the lower membrane surface were fixed with methanol for 10 min and stained with 0.1% Crystal Violetin 20% (v/v) methanol for 15 min, photographed and the cell number on each filter was counted in five randomly selected 200× fields.
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