Cell transient transfection

HCT116 cells were seeded for assays in 12-well plates at 300,000 cells/well and in 96-well plates at 10,000 cells/well. A549 cells were seeded for assays in 12-well plates at 200,000 cells/well and in 96-well plates at 5,000 cells/well. All transfection experiments were performed using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. The amount of plasmid and volume of Lipofectamine 2000 were optimized to achieve 70–80% transfection efficiency for all functional and molecular characterizations.