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. 2022 Nov 10;13:6814. doi: 10.1038/s41467-022-34562-5

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© The Author(s) 2022

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. 1 — a Chemical structure of A947. b Dose-response curves of A947 displacing a biotinylated SMARCA2/4-binding probe from recombinant SMARCA2 and SMARCA4 bromodomains. Data are presented as mean ± s.d. from 4 replicates. c Quantification of SMARCA2 and SMARCA4 protein levels by In Cell Western^TM following 20 h treatment of SW1573 cells. Data are normalized to DMSO control-treated cultures and presented as mean ± s.d. from 7 independent experiments. d Immunoblot analysis of respective proteins following 18 h treatment of SW1573 cells with a dose-response of A947. HDAC1 serves as a loading control. Data is representative of 2 independent experiments. e Licor-based quantification of SMARCA2 and SMARCA4 isoforms ectopically expressed in TOV112D cells following 24 h treatment with A947. Data is normalized to DMSO control treated cells. UniProt identifiers of the respective isoforms are shown in parentheses. f Pretreatment (1 h) of SW1573 cells with 20-fold molar excess of the respective inhibitors can block A947 (500 nM) -mediated degradation of SMARCA2. Total ubiquitin levels serve as a control for MLN-7243 and MG-132. HDAC1 serves as a loading control. g Immunoblots demonstrating FLAG-tagged SMARCA2 ortholog expression in human TOV112D and murine LA4 cells. Tubulin serves as a loading control. h Licor-based quantification of FLAG-tagged orthologs (human, mouse, rat) of SMARCA2 ectopically expressed in human (TOV112D) and murine (LA4) cell lines upon 24 h treatment with a dose-response of A947. Data are normalized to levels of the respective SMARCA2 ortholog in control (DMSO) lysates. Data in (f,g) were confirmed in 3 similar experiments. Source data are provided as a Source Data file.