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. 2023 Feb 13;24:25. doi: 10.1186/s13059-023-02867-3

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© The Author(s) 2023

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6. — H3.3 promotes binding of de novo expressed TFs. A–C Immunoblot of whole cell lysates from H3f3a^fl/fl;H3f3b^fl/fl;GtROSA26^CRE-ERT2 ESCs treated with either EtOH or 2 μM 4-Hydroxytamoxifen for 48 h followed by treatment with either DMSO or 0.5 μg/ml doxycycline for 48 h showing expression levels of H3.3 and Flag-tagged A BHLHA15, B HAND2, or C TWIST2. *Non-specific band ~30 kDa observed in all lysates. Total histone H3 or β-tubulin are used as loading controls. D–F CUT&Tag average profiles of D BHLHA15-Flag, E HAND2-Flag, and F TWIST2-Flag at promoter-proximal regions enriched in WT ESCs in WT and H3.3 KO ESCs. Three kilobases around the center of promoters are displayed for each analysis, and the number of regions profiled is indicated