Establishing and Treating Cell Lines

Human HT-1080 fibrosarcoma, A431 epidermoid carcinoma, MCF-7 breast cancer, and human embryonic kidney (HEK) 293T cells were obtained from the American Type Culture Collection and cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing glucose (4.5 g/liter), l-glutamine, and sodium pyruvate (11995073, Gibco). Medium was supplemented with 10% heat-inactivated FBS (16140071, Gibco) and 1% penicillin-streptomycin (10,000 U/ml; 15140122, Gibco). Cells were maintained at 37°C with 5% CO₂. A431 cells expressing GFP-E-cad were a gift from V. Bruntons (EH4 2XR, University of Edinburgh, UK) (63 (link)). In select experiments, cells were treated with the following pharmacological agents: Y-27632 (20 μM; Y0503, Sigma-Aldrich), ionomycin (0.5 and 1 μM; I0634, Sigma-Aldrich), blebbistatin (50 μM; B0560, Sigma-Aldrich), Rho inhibitor I (1 μg/ml; CT04-A, Cytoskeleton Inc.), LPA (50 μM; L7260, Sigma-Aldrich), paclitaxel (taxol equivalent) (1 μM; P3456, Thermo Fisher Scientific), colchicine (125 μM; C9754, Sigma-Aldrich), and importazole (50 μM; SML0341, Sigma-Aldrich).