Thymidine uptake in cells transduced with the MeiIRG retroviral vector.
BALB/3T3 A31 cells were transduced with MeiIRG retroviral vector, expressing the human nitrobenzylmercaptopurine riboside (NBMPR)-insensitive equilibrative nucleoside transporter eigene and a green fluorescent protein (GFP) reporter gene. Uptake of3H thymidine (Thd; 1.0 μmol/L at 4 μCi/mL) was measured in untransduced and MeiIRG-transduced A31 cells in the presence or absence of 100 nmol/L NBMPR or 10 μmol/L dipyridamole (DIP) in sodium-free MGA transport buffer. Cells were preincubated for 5 minutes at 22°C with either MGA buffer alone or MGA containing 100 nmol/L NBMPR or 10 mmol/L DIP before the addition of 3H Thd. Nucleoside uptake was determined at a constant time interval of 10 seconds. The values presented are from assays conducted in triplicate.
