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. 2022 Dec 19;7:392. doi: 10.1038/s41392-022-01247-w

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Characterization of optimized bat intestinal organoids and detection of the basal expression of antiviral genes. a Photomicrographs of optimized bat intestinal organoids on day 7 after passaging (magnification ×100). Scale bar, 100 μm. b Transmission electron microscopy illustrates the ultrastructural morphology of absorptive enterocyte (E), Paneath cell (P), goblet cell (G), and enteroendocrine cell (EE) in bat intestinal organoids. c Bat intestinal organoids were fixed and immunostained to label Villin + (green) enterocyte and MUC2 + (green) goblet cell. Nuclei and actin filaments were counterstained with DAPI (blue) and Phalloidin-647 (purple), respectively. Scale bar, 20 µm. d Phylogenetic analysis based on an alignment of horseshoe bat (Rhinolophus sinicus) IFNL1 and IFNL3 ORF cDNAs with other Chiroptera species and mammals. Bootstrap values (%) are indicated on each branch, and the scale for branch length is shown at the bottom of the tree. e, f GAPDH-normalized expression levels of IFN genes (e) and ISGs (f) in bat and human intestinal organoids. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. Two-tailed unpaired Student’s t test