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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. 6 — Silencing of IGFBP3 in tip-derived organoids induces alveolar-like differentiation. a IGFBP3 mRNA levels in tip-derived organoids after siRNA silencing (n = 6 per group). b Time-lapse images of control and IGFBP3 silenced tip-derived organoids during 25 h after transfection. c Representative images of Control and IGFBP3 silenced tip-derived organoids at 48 h post-transfection showed differentiation in alveolar-like structures in the IGFBP3 silenced group. d Expression analysis of stem cell markers in control and IGFBP3 siRNA-transfected organoids (n = 4 per group). e Expression analysis of AT2 cell markers in control and IGFBP3 siRNA-transfected organoids (n = 4 per group). f Expression analysis of AT1 cell markers in control and IGFBP3 siRNA-transfected organoids (n = 4 per group). g Immunofluorescence analysis of SFTPC and E-CAD in control and IGFBP3 silenced organoids. h Hematoxylin staining of tissue sections of lung explants from 10 pcw embryos cultured for 48 h with or without IGFBP3 peptide. A distinctive epithelial morphology can be observed between both groups: the control group showed a cubic epithelium (black arrows) while the IGFBP3 group remained in a more undifferentiated stage (red arrows). i Representative images of one of the 3 independent replicates of TGFβ protein arrays showing differences in p-SMAD1, p-SMAD2 and p-SMAD5 between control and IGFBP3 siRNA-transfected organoids. j Protein quantification in 3 independent replicates of TGFβ related phosphorylated proteins using the protein Arrays in control and IGFBP3 siRNA-transfected organoids (n = 3 per group). k Schematic representation of the effect of IGFBP3 silencing in tip-derived organoid on TGFβ and BMP signaling. *p < 0.05; **p < 0.01; ***p < 0.001