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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. Cells were collected for WCE preparation at the indicated time points. (A) Western blot analysis of HeLa WCEs and immunoprecipitated Flag-YY1 at the indicated time points. WCEs were probed with anti-Plk1, anti-Cyclin B1, and anti-YY1 antibodies. Immunoprecipitated Flag-YY1 was probed with anti-pT39 and anti-YY1 antibodies. (B) In vitro kinase assay using the same WCEs tested in (A) at the indicated time points with GST-YY1 attached to beads, in the presence of phosphatase inhibitors. Reactions were separated on SDS-PAGE, transferred to nitrocellulose membrane, and probed with anti-pT39, then anti-YY1 antibody.