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Cells were harvested 24 hrs later and cell extracts were assayed for luciferase, β-galactosidase and renilla luciferases, as stated in Table 2.
Cells were lysed 24-36 h later, and cell extracts were used to measure firefly luciferase and β-galactosidase activities, respectively.
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Genistein was added 8 hours later and cells incubated with compound for 16 hours.
Culture supernatants were collected 36 hours later and cell-free samples were stored at -30°C until cytokine assay.
The medium was removed 2 h later, and cells were lysed with 100 μl of DMSO.
Infectious medium was changed 12 h later and cells were kept in culture until following analysis.
Supernatants were removed 18 hours later and cells lysed for RNA isolation.
Transfection medium was discarded 10 h later and cells were washed twice using phenol red-free DMEM.
Culture medium was replaced with fresh medium 24 h later, and cells were irradiated with 5 Gy X-rays.
The medium was replaced 8 hours later and cells were collected for the next experiments 48 hours post-transfection.
Media was changed 4 hours later and cells were harvested 48 hrs after transfection for RNA isolation.
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