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. 2014 Jul 10;12:39. doi: 10.1186/s12964-014-0039-9

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Copyright © 2014 Ju 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/4.0), whichpermits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public DomainDedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Gαi does not interact with Necdin. (A) Schematic presentation of FLAG-tagged Gαo, Gαi, and chimeric proteins, Gαi/o and Gαo/i. A region of the GTPase domain of Gαo was replaced with the corresponding region of Gαi and vice versa. (B) Lysates of 293T cells expressing plasmids encoding 3 μg of each of the various FLAG-Gα constructs together with 10 μg of HA-Necdin were subjected to the immunoprecipitation assay with anti-Necdin antibodies, and the beads analyzed with antibodies against FLAG and Necdin. Input was loaded with 10% of 293T cell extracts used for the immunoprecipitation assay. The numbers beside blot indicate size marker (kDa).