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14C-uridine was added to the tubes. After gentle mixing the tubes were incubated for another two hours under identical conditions. The reaction was stopped by adding 1 mL of ice cooled 10% (w/v) trichloroacetic acid. The cells were separated from the medium using a glass fiber filter (1.2 μm pore size, Whatman GF/C). The filter was washed with ice cold solution of 5% (w/v) trichloroacetic acid and then transferred into a scintillation vial. After adding 2.5 mL of scintillation cocktail (Rotiszint® eco plus, Roth) the samples were measured using a Beckman Coulter LS 6500 scintillation counter. For positive control, a specific inhibitor of each process was added: replication was inhibited by doxorubicin (0.05-0.47 μM), translation by cycloheximide (3.5-178 μM), and transcription by actinomycin D (0.4-4.0 μM).
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