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. 2024 Aug 26;15:263. doi: 10.1186/s13287-024-03883-1

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 5 — IGFBP3 overexpression in tips and in tip-derived organoids becomes downregulated with the differentiation process. a Tip and stalk isolation after treatment of the bronchial tree with dispase and mechanical separation of the mesenchyme from the epithelium under the stereoscopic microscope. b Quantification of MUC5AC, c IGFBP3 and d miR-34a mRNA levels in tip and stalk enriched populations (n = 9 per group). e Immunofluorescence analysis of IGFBP3 and E-CAD in tip-derived organoids. f Microscope images showing gross morphology of tip-derived organoids cultured with control media vs differentiated organoids after 30-day culture with differentiation media (Pneumacult). g Immunofluorescence analysis of IGFBP3 and MUC5Ac in organoids cultured with control media and after 30-day differentiation with Pneumacult media. h Analysis of MUC5AC, i IGFBP3 and j miR-34a mRNA levels in control and differentiated organoids (n = 3 per group). k UMAP visualization of single-cell RNA sequencing data from control and alveolar differentiated tip-derived organoids, with colors representing the cell of origin. l UMAP visualization showing the different cell types identified in the control and alveolar differentiated tip-derived organoids. m UMAP illustrating IGFBP3 transcript expression in the control and alveolar differentiated tip-derived organoids