Cell culture and Transfection

A549 and H1299 cells were routinely cultured in Ham's F12 Medium or RPMI-1640 Medium containing 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere with 5% CO2. For plasmid transfection, cells were inoculated into 6-well plates at a cell density of about 70-80% and transfected after adherence to the wall. The reagents for transfection of plasmids were Lipofectamine 3000 (Promega) and were used according to the manufacturer's instructions. Cell culture medium was replaced after 4-6 h, and the transfected cells were collected after 24-48 h.