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. 2010 May 20;285(30):23105–23114. doi: 10.1074/jbc.M110.124990

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Transient Nutlin treatment promotes a prolonged 4N arrest in A549 cells. U2OS (A) and A549 (B) cells were treated with 10 μm Nutlin for 24 h. BrdUrd (20 μm) was added 6 h after the Nutlin addition. At the 24-h time point, cells were rinsed with phosphate-buffered saline and refed with normal medium (−Nutlin and BrdUrd). Cells were harvested at the indicated time points and analyzed by flow cytometry. The top panels for each cell line (labeled PI) show the DNA profile for the total population determined by propidium iodide staining. Middle panels (labeled BrdU) show the distribution of BrdUrd signaling for the whole population. BrdUrd-positive cells were gated, shown by the boxed regions in the middle panels. Bottom panels (labeled BrdU pos) show the DNA profile of BrdUrd-positive cells as gated in the middle panels. The numbers represent the percentage of BrdUrd-positive cells with 4N DNA content at each time point. The numbers are the average from three independent experiments, and the S.E. is shown in parentheses.