293T cells were transfected via calcium phosphate transfection method66. Briefly, 2.5 × 106 cells were seeded on 100 × 20 mm tissue culture grade plates (Corning) followed by transfection with 10–20 μg of plasmid DNA. Cells were harvested 48 h later by trypsin and neutralization with media, followed by centrifugation of the cells at 700 × g for 5 min followed by subsequent PBS wash and a second centrifugation step.
