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. 2017 Jun 20;7:3879. doi: 10.1038/s41598-017-04223-5

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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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TIP60 binds to the promoter of BRCA1 and FANCD2. The ChIP experiments were performed in HONE6 and TIP60-deficient (shTIP60) cells with anti-RNA Pol II (Pol II, positive control), mouse IgG (negative control), anti-TIP60 and anti-H4ac antibodies. The immunoprecipitated DNA fragments were quantified with qPCR with specific primers located at the transcription start site (TSS) of GAPDH, and at the TSS sites and at the 2 kb sites upstream (TSS up) or downstream (TSS down) from the TSS sites of BRCA1 and FANCD2. The protein enrichment at the TSS, TSS up, and TSS down sites were determined by the amount of pulldown DNA fragments relative to the amount of the input DNA. (A) RNA Pol II was enriched at the TSS sites of GAPDH. (B) TIP60 was enriched at the TSS sites of BRCA1 and FANCD2. (C) the enrichment of TIP60 was dramatically reduced in the TIP60-depleted cells. (D) H4ac was enriched at the TSS sites of BRCA1 and FANCD2. Depletion of TIP60 dramatically reduces the enrichment of H4ac at these sites. Each value represents the mean ± standard deviation from at least three experiments.