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. 2020 Jan 8;11:111. doi: 10.1038/s41467-019-13850-7

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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 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. 3 — a Scopolamine-treated Dclk1-DTR-ZSgreen-positive samples (n = 4 mice) show spatial separation from sham treated samples (n = 4 samples) along PC1 employing principal component analysis (PCA). b Top 25 differentially expressed genes (DEG) between the indicated groups, scale bar indicates log2-fold changes of genes (Z-Scores; positive lgFC indicates higher gene expression in the respective group). c Gene set enrichment analysis (GSEA) indicates negative correlation to previously described tuft-1 (neuronal) and tuft-2 (immune) phenotypes following scopolamine treatment; ES enrichment score. d Instead, tuft cells strongly enrich for enteroendocrine cell lineage signatures. e GSEA analysis of published enteroendocrine signature gene sets shows the adoption of an enteroendocrine progenitor subtype as well as increased differentiation into early and late EC cells, L cells and N cells of expanding tuft cells^13,32 (see Supplementary Data 1 for leading edge gene signatures); the enrichment for enterocyte signatures further confirms increased differentiation. EEC enteroendocrine cell, EC enterochromaffin cell, NES normalized enrichment score. f Master regulator analysis shows significant changes in transcription factor activity after scopolamine treatment, such as activation of Srebf2 and inactivation of Nme2, respectively (Z-Scores; Srebf2 p = 0.00086; Hnf4a p = 0.0014; Ewsr1 p = 0.0019; Taf9 p = 0.0019; Nme2 p = 0.0015; Sarnp p = 0.0011; Scand1 p = 0.00012). FDR false discovery rate, Act differential activity (red indicating increased, blue indicating decreased), Exp differential gene expression.