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. 2019 Feb 13;10:733. doi: 10.1038/s41467-019-08554-x

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© The Author(s) 2019

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. 8 — Identification of BRD9 as a potential therapeutic target for cancer treatment. a Expression levels of BRD9 mRNA in nine cancer types in which BRD9 was focally amplified. Gray: normal controls; white: tumors without BRD9 amplification; red: tumors with BRD9 amplification. BRD9 copy number status in each tumor specimen was estimated by a GISTIC analysis. b Expression of BRD9 protein was retrieved from the Human Protein Atlas dataset. A typical immunohistochemical staining of each cancer type is presented. Scale bar indicates 100 μm. c The heatmap of the genes that were significantly, positively co-expressed with BRD9 in nine cancer types. Each tumor is represented in a column and each gene in a row. The RNA expression of BRD9 is presented as a bar graph under the heatmap. The genes were arranged from top to bottom in descending order of their correlation with BRD9. d The pathways over-represented for BRD9-associated genes at the pan-cancer and individual cancer levels, according to pre-ranked gene-set enrichment analysis. e The functional hubs of pathways associated with BRD9 expression as analyzed by pre-ranked gene set enrichment. f BRD9 was knocked down with two independent shRNAs in four different breast and ovarian cancer cell lines. Knockdown efficiency was confirmed by real-time RT-PCR. g Four-day growth curves of cancer cell lines transfected with control or BRD9-specific shRNAs. h, i Soft agar assays of MDA-MB-231 (h) and OVCAR8 (i) cells expressing control and BRD9-specific hairpins. Scale bar indicates 1 cm. Quantification of the number of colonies from the soft agar assays on MDA-MB-231 (h) and OVCAR8 (i) cells. j In vivo xenograft tumor growth curve of MDA-MB-231 (upper) and OVCAR8 (lower) cells expressing control and BRD9-specific hairpins. k Representative pictures of tumors derived from control and BRD9-specific hairpin-expressing cells. Scale bar indicates 1 cm. l In vivo tumor weight curves for MDA-MB-231 (upper) and OVCAR8 (lower) cells expressing control and BRD9-specific hairpins. m Knockdown efficiency of xenograft tumors was confirmed by real-time RT-PCR. *t-test p-value < 0.05