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. 2012 Jun 22;40(17):8325–8335. doi: 10.1093/nar/gks599

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© The Author(s) 2012. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Mms4 phosphorylation through the cell cycle is Cdc28 and Cdc5 dependent. (A) Mms4 phosphorylation depends on Cdc28. Scheme of the experiment as explained in the text. (B) cdc28-td P_GAL1–10-UBR1 HA-MMS4 cells were collected at the indicated time points, and the DNA content was determined by flow cytometry. (C) Immunoblot analysis of HA-Mms4, Myc-Ubr1 and HA-Cdc28-td during the course of the experiment. The phosphorylated form of Mms4 is indicated as ‘Mms4-P’. The asterisk indicates an unrelated band that cross-reacted with the 12CA5 antibody. A Pounceau S-stained membrane was used as a loading control. (D) Mms4 phosphorylation requires Cdc5 activity. Scheme of the experiment as explained in the text. (E) The cell cycle progression of cdc5-1 HA-MMS4 cells was monitored by flow cytometry. (F) Immunoblot analysis of HA-Mms4 during the course of the experiment.