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. 2018 Oct 23;38(10):1717–1733. doi: 10.1038/s41388-018-0554-z

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© The Author(s) 2018

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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Fig. 3 — Synchronous inhibition of MEK and bromodomain-containing proteins inhibits cell proliferation and induces cell death in colon cancer cell lines. a MEK inhibitor-resistant human colon cell lines, T84, SNUC2A, and LS123 or the normal colon epithelial cell line CCD841CoN were treated with 30 nmol/L trametinib or 1 µmol/L JQ1 for 72 h. Cell proliferation was determined by cell counting and expressed as a percentage of the cell number prior to treatment. Mean values are presented, ± standard error (n = 3). Statistical significance was determined using a one-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. b Cells were treated as in (a) and then analyzed for annexin V positivity by flow cytometry. The mean percentage of annexin V-positive cells relative to DMSO controls is shown, ± standard error (n = 3). c Cells were treated as in (a) and cell lysates were analyzed by western blotting for the indicated proteins. d T84, SNUC2A, and LS123 cells, or the normal colon epithelial cell line CCD841CoN were treated with 30 nmol/L trametinib, 1 µmol/L JQ1 or the combination of both compounds for 14 d and cell proliferation was assessed by colony formation assay. e T84, SNUC2A, and LS123 cells, or the normal colon epithelial cell line CCD841CoN were treated with a matrix of trametinib and JQ1 for 4 d, and cell proliferation was assessed by the CellTiter-Blue assay (decrease in proliferation is shown by a shift from blue to red). Synergy was determined by the Bliss independence model (the excess above bliss score is indicated, with red indicating synergy)