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Annexin V/propidium iodide (PI) staining and flow cytometry

HW Hong Wang
QY Qingqing Yuan
MN Minghui Niu
WZ Wenhui Zhang
LW Liping Wen
HF Hongyong Fu
FZ Fan Zhou
ZH Zuping He
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The apoptosis percentages of male germ cells of wild type and P63(+/−) mice were detected using the Annexin V-FITC/PI kit by flow cytometry pursuant to the manufacturer’s instruction (Biolegend, London, UK). Male germ cells were obtained from P63(+/−) mice and wild-type mice using two-step enzyme and followed by the differential plating and culture for 24 h. It was feasible to identify and quantify apoptotic cells by conjugating FITC to Annexin V using flow cytometry. The cells were simultaneously stained with Annexin V-FITC (green fluorescence) and the non-vital dye PI (red fluorescence), which allowed the discrimination of intact cells (FITC−PI−), early apoptotic (FITC+PI−) and late apoptotic or necrotic cells (FITC+PI+). The procedure was carried out according to the protocol as described previously77.

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