Nunc delta surface 96 well cell culture plate

This assay was also performed using the luciferase assay system kit (Promega). Cells were seeded into a Nunc delta surface 96-well cell culture plate (Thermo Fisher) at a density of 2 × 10⁴ cells/well in 200 μl of cell culture media and left to adhere overnight at 37°C in a 5% CO₂ humidified atmosphere. The medium was removed, the cells were washed twice with 200 μl of cold PBS, and the plate was cooled on ice. Then, 20 pg/cell of recombinant adenovirus fiber knob was added to each well in 200 μl of cold PBS, followed by incubation on ice in a 4°C cold room for 1 h. The medium was then removed, and luciferase transgene encoding replication- incompetent viruses were added to the necessary wells at the required titer in 200 μl of cold serum-free RPMI 1640, followed by incubation on ice in a 4°C cold room for 1 h. The virus-containing medium was then removed and replaced with complete cell culture medium, and the cells were incubated for a further 45 h under normal cell culture conditions. From this point forward, the assay is identical to the GFP and luciferase transduction assays.