The tumor cells cultured in medium alone or containing control-oligo were used as control groups.
Medium was aspirated and medium containing 1% FCS alone for controls, or containing cisplatin, gemcitabine or darbepoetin at incremental concentrations, was then added to wells in triplicate, and the incubation was continued for 48 h in an aerobic or hypoxic (1% O2) incubator (Series II CO2 Incubator, ThermaForma, OH, USA).
After release from S-phase synchrony into medium containing nocodazole, similar to controls, the cells lacking Rad21 accumulated in mitosis.
Cultures were exposed only to medium containing 1% DMSO (controls) or medium containing different concentrations of decoction dissolved in 1% DMSO (600 μg/ml -2400 μg/ml), and incubated for 24 h.
Solvent controls received medium containing 0.4% dimethyl sulfoxide.
The results from their own study indicate that they used subnormal conditions, since at 2.5 and 5 μg arginine/ml, in the absence of lysine in the medium, the viral titer was less than 50%% of the controls (DMEM medium, containing 146 μg lysine and 84 μg arginine per ml).
Media was replaced after 2 hr of infection (VSV/SINV-LUC) or 5 hr of transfection (p166-LUC) with either vehicle-containing medium (control) or medium containing 50 µM PYR-41 or 40 µM MG132.
The medium containing glucose was used as control.
In addition, the negative control contained only medium, while the positive control was medium containing 5% DMSO.
