The amount of radioactivity incorporated was determined by using a liquid scintillation beta counter (Beckman Instruments, Inc.Madrid, Spain).
The amount of radioactivity incorporated was determined by using a liquid scintillation beta counter (Beckman Instruments, Inc .. Results shown are the mean [3H]-thymidine incorporation of triplicate wells.
Radioactivity was determined in both fractions by using a liquid scintillation counter.
The cells were further incubated for 16 h, then harvested and H-TdR incorporation monitored by using a liquid scintillation counter.
One microcurie of [H] thymidine (ICN Pharmaceuticals) was added for the final 18 hours of culture, and radioactivity incorporation was quantified by using a Betaplate scintillation counter.
The nuclear and mitochondrial DNA fractions were extracted as described above and evaluated for radioactivity by using a liquid scintillation counter.
Cell cultures were prepared with 1 × 105 T cells/well and 5 × 104 BMDCs/well (the R/S ratio is 2 1) in a 96-well plate, incubated for 4 days in 5% CO2 at 37°C, pulsed with 0.5 μCi H-thymidine (H-TdR) for 16 hours, and then harvested and counted for radioactivity by using a beta scintillation counter (Packard Instruments, Chicago, IL, USA).
Radioactivity taken up by the cells was determined using a scintillation counter.
Reactions were started by the addition of ATP with [P]ATP and incubated for 15 min. Finally, reactions were analyzed by SDS-PAGE autoradiographyandy, and quantified using a scintillation counter.
