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. 2009 Jan 12;184(1):67–82. doi: 10.1083/jcb.200801009

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© 2009 Zhang et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 6. — Overexpression of CDK6 and CDC25A accelerates S-phase completion. (A) Quantitative RT-PCR for endogenous (left bars) and total (right bars) expression of CDK6 and CDC25A in the H9 hESC line. The data represent the mean ± SEM (error bars) from three independent experiments. The value for the control clone was set to 1, and all other values were calculated with respect to this. (B) Overexpression of CDC25A and CDC25A shown by Western blotting. GAPDH is used as a loading control (a representative example from the H9 line is shown). Molecular masses are indicated in kilodaltons. (C) Increase in CDK6 kinase activity and CDC25A phosphatase activity in CDK6- and CDC25A-overexpressing hESC sublines, respectively. The value for the control subline was set to 100%, and all other values were calculated with respect to this. The data represent the mean ± SEM from three experiments performed in the H9 cell line. (D) Chart representation of the fraction of cells in S phase over time in CDK6- and CDC25A-overexpressing hESC sublines assessed by propidium iodide staining (a representative example from the H9 subline is shown). (E) Flow cytometry images showing cell cycle distribution in H9 control as well as H9 CDK6 and H9 CDC25A after transfection of control siRNA and NANOG siRNA and synchronization by nocodazole for 18 h assessed by propidium iodide staining. This figure represents an example of at least two independent experiments performed in the H9 subline.