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. 2017 Aug 2;7:7116. doi: 10.1038/s41598-017-07559-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Kaiso binds to the promoters of BCL2, BAX, and BIK. DNA fragments (200–500 bp) were obtained from nuclear lysates of HUVECs (A) and HMEC-1s (B), respectively, and were subjected to chromatin immunoprecipitation. Mouse monoclonal anti-Kaiso 6F/6F8-CHIP grade antibody was used for Kaiso/DNA complex precipitation and the precipitated DNA fragments were amplified using primers targeting promoter regions of BCL2 (−3902 to −1 bp) (A,a and B,a), BAX (−971 to −1 bp) (A,b and B,b) and BIK (−3000 to −1 bp) (A,c and B,c). Primer positions (horizontal arrows) are shown in the schematic drawings in the upper panel in A and B. Asterisks mark the positively amplified promoter regions. Normal mouse IgG was used as a negative control. M: DNA marker. (A) In HUVECs, Kaiso binds to the promoter regions −1201 to −1 bp of BCL2, −645 to −316 bp of BAX and −1201 to −1 bp of BIK (Asterisks). (B) In HMEC-1s, Kaiso binds to the promoter regions −301 to −1 bp of BCL2, −645 to −316 bp of BAX and −601 to −1 bp of BIK (Asterisks). Detailed primer positions and sequences were listed in Supplementary Table S2.