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. 2020 Mar 12;43(3):431–444. doi: 10.1007/s13402-020-00497-6

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — JQ1 affects stemness-related functions and downregulates the expression of stemness-related genes. a. Gene Set Enrichment Analysis (GSEA) identifying stemness-related biological functions. b. Manual screening uncovering 13 stemness-related genes. Heat map indicating fold changes of JQ1-deregulated genes compared to those in non-treated cells (incubated with vehicle). c-g. JQ1 effect on mRNA levels of the stemness-related genes identified in b corroborated by qRT-PCR. MDA-MB-231 (c-f) or BT549 (g) cells were treated with the indicated doses of JQ1 (24 h) after which qRT-PCR was performed for the indicated genes. c and g. CD44 and CD24 mRNA levels were used to calculate the CD44/CD24 ratios in JQ1 exposed cells compared to control cells. d and gALDH1A1 mRNA levels were significantly downregulated in cells treated with JQ1 at both doses. e. JQ1 significantly decreased ABCG2, ITGA6, MYC and EPCAM levels in MDA-MB-231 cells, and modestly affected the EZH2 level. f.GJA1, PIAS3, RUNX2 and SOX9 mRNA levels were significantly altered by JQ1, while just a slight decrease was found for FSTL1. The qRT-PCR conditions are described in material and methods. The results shown are averages of 3 independent experiments performed in triplicate. Student t-test: * p < 0,05, ** p < 0,01 and *** p < 0,001