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. 2012 May 4;7(5):e36372. doi: 10.1371/journal.pone.0036372

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Ito 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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(A) U2OS cells expressing mKO2-CycinB1 were treated with siRNAs and time lapse images were recorded by LCV100. The time (hr) between the first appearance of cytosolic mKO2-CyclinB1 signal and its translocation into the nucleus was measured in the time lapse images of each cell population. In Cdc7-depleted U2OS, CyclinB1 does not accumulate in cytoplasm. However, co-depletion of Cdc7 and p53 caused CyclinB1 accumulation in cytoplasm for a longer period. The P-values of the two-tailed unpaired t-test were calculated by Prism software. (B) Western analysis of the whole cell extracts of U2OS cells treated with indicated siRNAs for 48 hrs. A phosgel was used for the detection of MK2. Other proteins were detected on a 4–12% gradient gel.