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Cell cycle analysis by DNA content

QW Qiong Wang
GH Guopei He
MH Mengmeng Hou
LC Liutao Chen
SC Shangwu Chen
AX Anlong Xu
YF Yonggui Fu
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Cells were synchronized by starvation in DMEM supplemented with 1% FBS for 72 h and then released into the cell cycle by stimulation with 10% FBS for 6 h before cell cycle analysis. Cells were harvested with trypsinization and fixed in 70% ethanol overnight at −20 °C. After washing with phosphate-buffered saline (PBS), the cells were incubated with 0.2 mg/ml DNase-free RNase A (Sigma), 20 μg/ml propidium iodide (Sigma) and 0.1% v/v Triton X-100 (Sigma) in PBS for 30 min on ice in the dark. Cell data were collected with a FACS Calibur flow cytometer (BD Biosciences) and analyzed by Modfit software.

Copyright and License information: The Author(s) ©2018
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