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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 specific KBSs on BCL2, BAX and BIK promoter. Nonradioactive EMSA was performed using probes targeted to the promoter positions that contain putative KBSs (BCL2: −193 bp, BAX: −389 bp, BIK: −75 bp, −36 bp). Wild type probes were labeled with 6-FAM and were incubated with cell nuclear extracts from HUVECs overexpressing Kaiso (A, lane 2–5) or with recombinant protein GST-Kaiso (B, lane 2–5). Labeled wild type probes (FAM-probe) alone served as a negative control (lane 1). Monoclonal mouse anti-Kaiso 6F/ 6F8-CHIP grade antibody (A, lane 6) and monoclonal mouse anti-GST antibody (B, lane 6) were used to detect super gel shift. Unlabeled wild type probes (Competitory-probe) served as competition. KBS-mutated probes (FAM-mut-probe and Competitory-mut-probe) served as a negative control and competition.