Proliferation and Migration Assays of BT474 Cells

BT474 cells were transfected and seeded into 96-well plates (6×10³ cells per well) for the proliferation activity which was determined at 0, 6, 12, 18, and 24 h post-transfection using the Cell Counting Kit-8 (Dojindo, Japan) according to the manufacturer’s instructions. For the migration assay, transfected BT474 cells were treated with mitomycin C (10μg/ml, R&D Systems, USA) for 10 h to prevent proliferation, and equal number of cells were then seeded into the 24-Well Millicell plates (Millipore, Germany) containing an 8-μm pore membrane as previously described [14 (link)] and the transwell-containing plates were incubated for 24 h.