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. 2017 May 10;7:1684. doi: 10.1038/s41598-017-01943-6

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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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Expression system for Brd4 mutants. (a) Schematic structure of recombinant full-length human Brd4 protein (Brd4-HA) and Brd4 fragments f1–f9. BD1/2 = bromodomain 1/2, ET = extra-terminal domain, PID = P-TEFb-interacting domain, HA = hemagglutinin tag, NLS = nuclear localization signal. (b) Expression levels of HA-tagged Brd4 mutants, assessed via western analysis of the HA tag. α-Tubulin served as loading control. Samples were diluted with loading buffer prior to loading to visualize all signals on a single membrane (dilution ratios: lanes 2 and 9, 1:1; lanes 1, 3, 4, 6, 10 and 11, 1:10; lanes 5, 7 and 8, 1:20). Raji-luc cells expressed an non-tagged luciferase and served as negative control.