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. 2011 Aug 29;31(15):1975–1987. doi: 10.1038/onc.2011.372

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Copyright © 2012 Macmillan Publishers Limited

This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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RASSF1A interacts with Cdc20 during mitosis. (a) RASSF1A was immunoprecipitated from asynchronized (AS), mitotic (M) or MG132-treated mitotic (M+MG132) HeLa cells as indicated. The immunoprecipitate and 2% of the input WCL was subjected to immunoblotting (IP, immunoprecipitation; WCL, whole-cell lysate). (b) Cdc20 interacts with multiple domains on RASSF1A. FLAG-Cdc20 and different fragments of HA-RASSF1A were co-transfected into HEK293 cells. The immunoprecipitate of FLAG-cdc20 was analyzed by immunoblotting. (c) D-box and KEN-box of RASSF1A are dispensable for the interaction with Cdc20. Different D-box deletion mutants of RASSF1A were co-transfected with FLAG-Cdc20 into HEK293 cells and the immunoprecipitation–immunoblotting assay was performed as in (b). (d) Mapping of D-box-dependent and -independent Cdc20-binding domains of RASSF1A. FLAG-Cdc20 and different fragments of HA-RASSF1A^DBD+KD were co-transfected into HEK293 cells and the immunoprecipitation–immunoblotting assay was performed as in (b).