Cell apoptosis assay was conducted based on a study method [44 (link)] with some changes. Apoptosis of control and BHMC-treated MCF-7 cell was evaluated using AnnexinV-FITC/PI apoptosis detection kit (Becton Dickinson, USA). In brief, harvested cells were washed and re-suspended in 100 µL of PBS. Then, the cell was stained with 10 µL of AnnexinV-FITC/PI solution and incubated for 20 min in dark prior to analysis with BD FACS Calibur flow cytometer (Becton Dickinson, USA). Approximately, 10,000 cells in the population were captured. The experiment was qualitatively repeated for three times.
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