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. 2013 Jul 25;8:10. doi: 10.1186/1747-1028-8-10

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Copyright © 2013 Hagen et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Downregulation of Cdk4 results in impaired phosphorylation of BAD (ser136) and upregulation of protein phosphatase type 2A (PP2A). Cells stably expressing pLKO.1 or shCDK4 were irradiated at 2 Gy. Protein lysates were then prepared at different times (0, 0.5, 1 & 2 hours) post-irradiation and were immunoblotted with anti-Bad(ser136), anti-Mcl-1(ser159/thr163) and anti-Bcl-2(ser70) antibodies, and their counterparts detecting total levels of those proteins. (B) The levels of PP2A and PP2B were further determined by immunoblotting with anti-PP2A and anti-PP2B antibodies. Cells transfected with pLKO.1 were used as control. The β-actin controls for (B) is the same as for (A).