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5-ethynyl-2’-deoxyuridine (EdU) staining

RJ Renbing Jiang
ZD Zhibing Dai
JW Junshen Wu
SJ Suzhi Ji
YS Yachao Sun
WY Wenpeng Yang
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After transfection, U2OS or HOS cells (1 × 104) were maintained in RPMI 1640 medium containing EdU (50 μM, RiboBio) at 37 °C for 4 h, treated with 4% PFA for 30 min, and maintained with glycine for 10 min. Then, these cells were treated with 0.3% Triton-X-100 solution for 15 min and rinsed with PBS three times. Next, each well was treated with 200 μL 1× Apollo reaction liquid for 20 min and the nuclei were stained with 4’,6-diamidino-2-phenylindole (5 μg/mL). The cells were imaged by a fluorescence microscope (Olympus) and the number of EdU-positive cells was calculated.

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