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. 2022 Mar 2;117(1):9. doi: 10.1007/s00395-022-00919-6

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© The Author(s) 2022

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/.

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Fig. 5 — mRNA expression analysis of HCF reveals repression of ECM and collagen function after treatment with lyco-s. a Out of 61,043 transcripts, 18,964 remained after exclusion of low-expressed transcripts. Volcano plots show over 4000 differentially expressed genes (DEGs) for all three treatments. Treatment with lyco-s led to a similar amount of significantly up- (red), but not down-regulated genes (blue) compared to bufalin similars. n = 3 biological replicates (donors). Discoveries (Q = 0.05) adjusted following BY. b Treatment with lyco-s leads to increased deregulation of central pathways related to ECM function, including ECM organisation (orange), collagen binding (green), or both (yellow). GO:BP analysis of DEGs highlights downregulation of ECM and myofibroblast functions (c) as well as metabolism (d) after treatment with lyco-s. In contrast, treatment with buf-s-18 led to downregulation of genes involved in cilia (e) and DNA damage responses (f). For samples treated with buf-s-14, no terms were significantly enriched after multiplicity adjustment. Dotted line indicates significance threshold (p_adj. < 0.05). g Treatment with lyco-s (purple) repressed most high to medium-expressed collagen isoforms in HCF. The top 5 collagens are among the top 20 overall genes (compare Supplementary Fig. 4d). Mean (bars) and single biological replicates (circles), asterisk indicates significance in Two-way ANOVA (p < 0.05), adjusted following Dunnett (including buf-s-14 + buf-s-18, see Supplementary Fig. 4d)