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. Author manuscript; available in PMC: 2009 Jul 25.
Published in final edited form as: Cell. 2008 Jul 25;134(2):256–267. doi: 10.1016/j.cell.2008.05.043

Figure 4. Cdh1 associates with Cdc14B in G2 in response to DNA damage.

Figure 4

(A) Cdc14B moves from the nucleolus to the nucleoplasm in response to DNA damage. U2OS cells transfected with a construct expressing GFP-tagged Cdc14B were synchronized as described in (2A) and then either treated with DRB [+/−caffeine (CAF)] or subjected to ionizing radiations (IR). Cells were then collected and analyzed by direct immunofluorescence. Left panels: micrographs of representative cells showing the subcellular localization of Cdc14B. Right panel: Quantification of cells with nuclear Cdc14B fluorescence at the indicated times post pulse with doxorubicin (n=3, ± SD).

(B) Cdc14B translocation to the nucleus is independent of ATM. The experiment was performed as in (A), except that asynchronous ATM+/+ and ATM−/− fibroblasts were used (n=2).

(C) The experiment was performed as in (A), except that GFP-tagged Cdc14B(T426A) was used (n=3).

(D) Cdh1 binds to Cdc14B in a DNA damage-dependent manner. U2OS cells infected with either an empty retrovirus (EV) or viruses encoding FLAG-tagged Cdc14B were synchronized and treated with DRB as described in (A). Cells were collected two hours later, and whole cell extracts (WCE) were immunoprecipitated (IP) with anti-FLAG resin. The indicated proteins were detected by immunoblotting.