Transfection of HEK 293 T Cells
Human epithelial kidney 293 T (HEK 293 T) cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), penicillin (100 U/L), and streptomycin (100 mg/L) at 37 °C in a 5% CO2. One day before transfection, cells were transferred to 12-well culture plate to grow to approximately 70% to 80% confluence in an antibiotic free medium. Cells were then transfected with 2 μg plasmid DNA in each group of minigenes (empty pSPL3-control, pSPL3-WT and pSPL3-Mutation (MUT)) using OPTI-MEM® IMedium and Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.