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. 2024 Apr 11;43(10):1965–1989. doi: 10.1038/s44318-024-00086-5

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© The Author(s) 2024

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) RT-qPCR and Western blot examining TEAD2 expression in both 2iL- and SL-ESCs. mRNA expression was tested in triplicate in three independent experiments. Data are presented as the mean ± SD. P-values were determined using two-sided Student’s t-test (***p < 0.001). Quantification of protein signal was performed using Fiji image analysis software. (B) Strategy of generating Tead2-knockout cell lines in SL-ESCs. (C) Representative cellular morphologies of wild-type ESCs and two clones of Tead2^−/− and Tead2^+/− ESCs during the SL-to-2iL transition at day 0, 3, and 6. Scale bar, 100 μm. (D) PCA of the RNA-seq data from wild-type ESCs and Tead2-knockout ESCs collected at different time points during the SL-to-2iL transition. (E) Heatmap of the expression of 2i- and serum-specific genes during the SL-to-2iL transition with or without Tead2-knockout. Clustering analysis and the enrichment of GO terms in each group of genes are also provided. The enrichment p-value was calculated by using Metascape software. (F, G) RT-qPCR analysis testing the expression of serum-specific genes (F) and 2i-specific genes (G) in wild-type and Tead2-knockout cells on day 0 and day 6 of the transition. Data are presented as the mean ± SD. Indicated significances are testing using Student’s t-test analyses (*p < 0.05, **p < 0.01, ***p < 0.001). n = 3 biological replicates. Source data are available online for this figure.