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. 2023 Sep 11;13:14960. doi: 10.1038/s41598-023-41951-3

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© The Author(s) 2023, corrected publication 2023

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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SGG UCN34 induces pre-cancerous transformation of murine colon tissue. (a) Immunofluorescence analysis of Pdpn (green) and CD34 (red) in sections of colonic tumor-free regions of mice colonized with SGM (top) or SGG UCN34 (bottom) (DAPI/nuclei in blue and E-cadherin in grey). Scale bar 50 µm. Images were acquired with SP8 microscope (Leica) and a ×40 objective. Graph presents the percentage of colocalization of Pdpn and CD34 signals per field of view (FOV). The quantification was done by Imaris (version8) software on SGM (n = 8) and SGG UCN34 (n = 7) colonized mice, using at least 6 images per mouse and showing 1.1% vs 3.1% of Pdpn + CD34 + signal respectively (*t-test p = 0.01). (b) Experimental design of ex vivo organoid formation originating from macroscopically tumor-free sections of colon tissue colonized by SGG UCN34 or SGM over 12 weeks in the AOM-induced CRC model. (c) The number of organoids with normal or cystic morphology defined as colonospheres or colonoids with a central empty lumen and polarized epithelium (actin staining). (d) The number of organoids with compact morphology defined as colonospheres or colonoids with a lumen full of cells and depolarized, restructured actin filaments. (e). Microscopic pictures showing DAPI/nuclei labeling (blue), Phalloidin/actin (green) and Ki67/proliferation marker (red). Images were acquired with Opterra Multipoint Scanning Confocal microscope (Bruker) at ×63 objective and analyzed by Fiji software. Scale bar 50 µm.