HeLa cells were seeded in 6-well plates at a density of 1 × 106/well and treated with either 0.1% DMSO or alloimperatorin (50, 100, and 150 µM), respectively, for 48 hours. The cells were collected and fixed with 70% ethanol overnight. They were combined with RNase A at 37°C under dark conditions. The incubation was performed with 1 ml PI solution (20 μg/ml 1% Triton X-100 in PBS) for 30 minutes. CellQuest software (BDIS) was used to evaluate the cell cycle by flow cytometry (BD Bioscience, MA, USA).

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