SV40 DNA Transfection and Lytic Induction

iD98/HR1 cells were plated on coverslips in 6-well plates and treated with 100 μg/mL of PAA, 24 h prior to being mock transfected or transfected with isolated SV40 DNA. SV40 DNA isolation was carried out as previously described (64 (link)). The transfection mix was incubated with the cells for 4 h in the 37°C cell incubator (humidified, with 5% CO₂), and then the cells were washed with 2 mL of DMEM with 10% FBS (D10F) before another 2 mL of D10F was added as growth medium. The cells were grown for 1 h, and then 100 μg/mL PAA was added back to the medium. Cells were grown for another 2 h and then were induced to enter EBV’s lytic phase by the addition of 200 nM 4-OHT. At 48 h after transfection, the cells were incubated with 10 μM EdU for 1 h and then fixed for click chemistry and immunofluorescence assays.