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. 2018 Jan 4;10:1. doi: 10.1186/s13148-017-0434-3

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 6 — Changes in histone modifications elicited by combination treatment. a Changes in H3 and H4 acetylation were detected after single and combined treatment in VM-Cub1 and UM-UC-3 cells by acid histone extraction and Western blot analysis. Total histone 3 served as a loading control. b Chromatin immunoprecipitation analysis of H3K27 acetylation at transcriptional start sites (TSS) as well as 2 kb upstream (− 2 kb) and 2 kb downstream (+2 kb) of BCL-2, BCL2L1/BCL-X, BIRC5/Survivin, c-MYC, CDKN1C/p57 and SKP2 genes after single and combined treatment of VM-Cub1 cells. Immunoprecipitated DNA amounts measured by qPCR for each condition are expressed as percentage of input DNA. Additionally, H3K4 trimethylation was analyzed at the TSS of the same genes. Rabbit IgG served as a background control. Differences between samples with combination treatment and DMSO treated cells were analyzed using Student’s t test (***p ≤ 0.001, **p ≤ 0.01, *p ≤ 0.05)