Paneth cells inhibit intestinal stem cell proliferation through the bone morphogenic protein 7 pathway under rotavirus-mediated intestinal injury

Figure 1 Rotavirus infection has been detected in Paneth cells. A: Schematic representation of genome sequencing of the rotavirus (RV) enteric cytopathogenic human orphan virus strain; B: Uniform manifold approximation and projection of intestinal epithelial cells from uninfected and infected mice; C: Expression of representative genes in clusters from B; D: Distribution of cells containing RV genes; E: Number of RV genes per RV⁺ cell grouped by cell type; F: Comparison of the proportions of Paneth cells in uninfected and infected mice; G: C57BL/6J mice aged 5 weeks were orally gavaged with the RV enteric cytopathogenic human orphan virus strain and sacrificed 72 hours post-infection; H: RV load in uninfected and infected mice (n = 6, mean ± SD). Viral RNA levels were measured by reverse transcription quantitative polymerase chain reaction and normalized to glyceraldehyde-3-phosphate dehydrogenase; I: Immunofluorescence staining of Paneth cells (lysozyme 1, red) in uninfected and infected mice. Nuclei were counterstained with 4’,6-diamidino-2-phenylindole (blue). Scale bar = 50 μm; J: Quantification of lysozyme 1 fluorescence intensity in Paneth cells from uninfected and infected mice. Fluorescence intensity was normalized to the uninfected group (set to 1); K: Flow cytometric analysis of Paneth cells in uninfected and infected mice. Representative fluorescence-activated cell sorting plots showing the percentage of Paneth cells in both conditions; L: RV load in sorted Paneth cells from uninfected and infected mice (n = 6, mean ± SD), measured by quantitative real-time polymerase chain reaction and normalized to glyceraldehyde-3-phosphate dehydrogenase. Statistical significance was determined using the non-parametric Mann-Whitney test between groups. ^bP < 0.01. EC: Enteric cytopathogenic human orphan virus; UMAP: Uniform manifold approximation and projection; EEC: Enteroendocrine cell; TA: Transit-amplifying cell; Olfm: Olfactomedin; Mki67: Marker of proliferation Ki-67; Lyz: Lysozyme; RV: Rotavirus; DAPI: 4’,6-diamidino-2-phenylindole; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.