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Analysis of cell apoptosis and cell cycle arrest by flow cytometry

SY Shuangzhi Yuan
YS Yong Sun
WC Wenqiang Chang
JZ Jiaozhen Zhang
JS Jifa Sang
JZ Jiachun Zhao
MS Minghui Song
YQ Yanan Qiao
CZ Chunyang Zhang
MZ Mingzhu Zhu
YT Yajie Tang
HL Hongxiang Lou
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Cell apoptosis and cell cycle arrest were analysed by flow cytometry according to the literature66–68 with minor modifications as followed. Briefly, after overnight incubation, the cells were exposed to 5% serum medium with concentrations of 0, 2.5, 5, and 10 μM DPL (24) for 24 h and then harvested and washed with PBS. After the supernatant was removed by centrifugation, the cells were resuspended in 400 μL of binding buffer and incubated at room temperature in the dark for 15 min with 5 μL of Annexin V-FITC, followed by 5 μL of PI (50 mg/L) for another 5 min. The apoptotic ratio was analysed by flow cytometry (Becton Dickinson, USA) using WinMDI 2.9 software. HUVECs were treated with 0, 1.25, 2.5, or 5 μM DPL (24) for 24 h, and cell cycle arrest was detected by flow cytometry analysis coupled with PI staining.

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