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Flow cytometry for cell cycle and apoptosis

RL Ruijie Liu
PD Ping Deng
YZ Yonglian Zhang
YW Yonglan Wang
CP Cuiping Peng
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Tali® Cell Cycle Kit (Invitrogen) was used for cell cycle detection. Briefly, 4 × 106 cells were washed by Dulbecco’s phosphate-buffered saline (DPBS; Gibco) and cells were fixated by ice-cold 70% ethanol (Sigma-Aldrich, St. Louis, MO, USA) in distilled water at − 20 °C overnight. In the dark, cells were stained with Tali® Cell Cycle Solution that was composed of propidium iodide (PI), RNase A, and Triton X-100 for 30 min. Eventually, cell proportion of each phase (G0/G1, S, and G2/M) was respectively determined using the flow cytometer (BD Biosciences, San Diego, CA, USA).

Cell apoptosis was detected by ApoDETECT Annexin V-FITC Kit (Invitrogen). Cell density was adjusted to 4 × 105 cells/mL in 1× binding buffer, and 10 μL Annexin V-FITC was added to 190 μL cell suspension. After incubation for 10 min at room temperature, cells were incubated with 10 μL PI stock solution. Cell analysis was performed by a flow cytometer (BD Biosciences). The apoptotic rate (%) was calculated using the ratio of apoptotic cells and total cells.

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