The invasion ability of breast cancer cells was assessed in vitro by employing a transwell assay, as described in our previous study.
29 (link) In summary, total of 3 × 105 breast cancer cells (Hs578T or MDA‐MB‐231) with ME1 knockdown or a scrambled control were placed in a suspension containing 2% FBS. These cells were then seeded onto the upper chamber of Falcon transwells (Falcon, Corning, USA), which were coated with Matrigel (BD Biosciences, MA, USA) to facilitate the invasion assay. Subsequently, the cells were placed in a CO2 incubator at 37°C for either 12 or 24 h. After the incubation period, any remaining cells in the upper chamber were removed using cotton swabs, whereas cells on the undersurface of the transwells were fixed using a 10% formaldehyde solution. The cells were then stained with crystal violet solution, and the number of breast cancer cells was determined by counting the three fields with a phase‐contrast microscope. Each experiment was completed three times to ensure accuracy.
29 (link) In summary, total of 3 × 105 breast cancer cells (Hs578T or MDA‐MB‐231) with ME1 knockdown or a scrambled control were placed in a suspension containing 2% FBS. These cells were then seeded onto the upper chamber of Falcon transwells (Falcon, Corning, USA), which were coated with Matrigel (BD Biosciences, MA, USA) to facilitate the invasion assay. Subsequently, the cells were placed in a CO2 incubator at 37°C for either 12 or 24 h. After the incubation period, any remaining cells in the upper chamber were removed using cotton swabs, whereas cells on the undersurface of the transwells were fixed using a 10% formaldehyde solution. The cells were then stained with crystal violet solution, and the number of breast cancer cells was determined by counting the three fields with a phase‐contrast microscope. Each experiment was completed three times to ensure accuracy.
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