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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-deficient HONE6 cells are sensitive to cisplatin. (A) TIP60 was depleted by two specific shRNAs (shTIP60-1, and shTIP60-2) packed lentivirus in HONE6 cells. The shLacZ was used as a nontargeting control. The knockdown efficiency of TIP60 was determined by qRT-PCR and normalized by the level in the control cells. Each value represents the mean ± standard deviation from at least three experiments. (B) Cell lysates were subjected to Western blotting with an anti-TIP60 antibody. The total cell lysates and the nuclear fraction of cells were indicated. Full-length blot is presented in Supplementary Figure S5. (C and E) Cytotoxicity assay of TIP60-deficient cells. Cells were treated with various concentrations of cisplatin or MMC for 96 hours. Cytotoxicity was determined by MTT assay, with relative viability being normalized to colonies of no treatment control cells of each cell line. (D and F) The colony formation assay. Cells were chronically treated with cisplatin or MMC as indicated and incubated for 10 days. The resulting colonies were stained with 1% crystal violet. Relative viability was normalized to colonies of no treatment control cells of each cell line. Each value derived from the MTT assay and the colony formation assay represents the mean ± standard deviation from at least three experiments.