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. 2024 Oct 8;15:348. doi: 10.1186/s13287-024-03958-z

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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/.

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Fig. 3 — VIP modulates Lgr5-EGFP + progenitor cell number and progeny in vitro. A Experimental outline and treatment scheme. B Quantification of Lgr5-EGFP + progenitor cells in VIP-treated organoids by flow cytometry (n = 10 per group). C Analysis of cellular proliferation following VIP treatment by EdU assay (n = 4 per group). D Flow cytometry analysis of Lgr5-EGFP + cell progeny following VIP treatment (n = 6 per group). E, PCA of transformed count data from bulk RNA sequencing of Lgr5-EGFP + progenitor cells isolated from intestinal control organoids and organoids treated with VIP. F Hallmark gene-set enrichment analysis of differential gene expression in Lgr5-EGFP + progenitor cells isolated from VIP-treated organoids; Ctrl = control, NES = normalized enrichment score. G VIP-induced differential upregulation of hallmark pathways in Lgr5-EGFP + progenitor cells compared to treated whole organoids. *p < 0.05, **p < 0.01. Data are shown as means ± SEM, n = number of biological replicates. Statistical analysis was performed by paired Student’s t-test in (B–D)