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. 2014 Aug 28;5(8):e1389. doi: 10.1038/cddis.2014.346

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Copyright © 2014 Macmillan Publishers Limited

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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A pool of Len/VD3-induced Bik protein was free from Bcl-2 and Bcl-x_L. (a) IP assay of Bik in JEKO-1 cells. The Bik IP assay was performed in lysate of cells (10⁷ cells) cultured for 4 days with or without 1 μM Len and 100 nM VD3. The expression of Bik, Bcl-2, Bcl-x_L and Mcl-1 was analyzed in pellet (IP) and supernatant (OUT). (b) Reciprocal Bcl-2 and Bcl-x_L IP assays. IP assays were performed by the simultaneous addition of both anti-Bcl-2 and anti-Bcl-x_L antibodies to the lysates. The protein expression was analyzed in the total lysate (IN), pellet (IP) and supernatant (OUT) fractions