Fig. 5

Both canine jejunal tissues and enteroids express mRNA for markers of various epithelial cell lineages. a Representative RNA in situ hybridization (RNA-ISH) images reveal expression of stem cell markers LGR5, SOX9, and EPHB2 on both intact jejunal tissues and enteroids. SOX9 is also a marker of enteroendocrine and tuft cells, while EPHB2 is also a marker of Paneth-like cells. Arrows indicate representative positive red areas in crypts (black arrows) or villi (yellow arrows). Representative images from at least n = 15 organoids per condition. b Representative images of gene expression for markers of epithelial lineage, including FZD5 (Paneth-like cell), ALP (absorptive epithelium), and Neuro G3 (Enteroendocrine cells) in both intact jejunal tissues and enteroids, as determined by RNA-ISH. Representative images from at least n = 15 organoids per condition. c Semi-quantitative scoring of RNA-ISH staining (box and whisker plots) for expression of stem cell markers (LGR5, SOX9), Paneth-like cell markers (FZD5, EPHB2), absorptive epithelial markers (ALP), and enteroendocrine cells (Neuro G3) in both intact jejunal tissues and enteroids. Specific sites include Enteroid Crypt (Ent Cryp), Enteroid Villus (Ent Vill), whole tissue Jejunum Crypt Base (J Cry B), Jejunum Crypt Neck (J Cry N), Jejunum Villus (J Vill), and Jejunum Villus Tip (J V T). Scoring of at least n = 6 images per condition. d Semi-quantitative expression of Paneth cell markers IL-17, CBD 103, and CATH as well as tuft cell marker Dclk1, in both intact jejunal tissues and enteroids, in specific sites as above. Cells and tissue were counterstained with hematoxylin. Scoring of at least n = 6 images per condition