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. 2011 Jan 6;6(1):e15928. doi: 10.1371/journal.pone.0015928

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Rizkallah et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Stable HeLa-Flag-YY1 cells were synchronized by double-thymidine block and then released. (A) Analysis of the cell-cycle progression of HeLa cells released after double-thymidine block using fluorescence-activated cell sorting. Cells were stained with propidium iodide and analyzed based on their DNA content. An asynchronous population of cells was used as a control. (B) Whole cell extracts were prepared from HeLa-Flag-YY1 cells collected at the indicated times after release from double-thymidine block. Total WCE were analyzed on a Western blot after SDS-PAGE separation, and probed with anti-Plk1, anti-Cyclin B1, and anti-YY1 antibodies. Flag-YY1 was immunoprecipitated from the extracts of each time point, and then analyzed on a Western blot using anti-pT39 and anti-YY1 antibodies.