For the scratch assays, cells were seeded in triplicate and when they reached 95–100% confluence, were serum starved with 0.1% FBS-containing media for 12 h. Subsequently, a scratch was made across the cell layer using a P-200 pipette tip, and cell migration was monitored by recording images at indicated time points post-scratch. The area of the scratch was quantified using the MiToBo plug-in for ImageJ software and plotted as a percentage of total area.
For the transwell migration assay, cells were trypsinized and re-seeded in triplicate in migration chambers (BD Bioscience, Bedford, MA) in serum-free medium. 24 hours (MCF10AT1 cells) or 48 hours (MCF10CA1a cells) after cell seeding, the experiment was performed and results quantified as previously described [33 (link)].
For the transwell migration assay, cells were trypsinized and re-seeded in triplicate in migration chambers (BD Bioscience, Bedford, MA) in serum-free medium. 24 hours (MCF10AT1 cells) or 48 hours (MCF10CA1a cells) after cell seeding, the experiment was performed and results quantified as previously described [33 (link)].
