Supplementary MaterialsFIG?S1. pets given to inoculation prior, and fasted identifies pets separated from dams for 24 h s ahead of inoculation. (C) Preliminary body weights of baby rabbits inoculated with WT disease. (A and B) Hematoxylin-and-eosin-stained colonic parts of serious hemorrhage in lamina propria and colonic lumen from pets infected using the WT stress at 36 hpi. Arrowheads in -panel A reveal either a location of hemorrhage in the lamina propria or hemorrhage growing towards the lumen (inset in -panel A). (B) Hemorrhage and epithelial cell sloughing in colonic lumen. Size pub, 100 m. Download FIG?S2, TIF document, 2.1 MB. Copyright ? 2020 Kuehl et al. This article can be distributed beneath the conditions of the Innovative Commons Attribution 4.0 International permit. FIG?S3. Selection of IL-8 manifestation in colons of infected infant rabbits. (Top left, plot) Percentage of IL-8-expressing cells in each ST6GAL1 field of view from colonic tissue sections stained with probe to rabbit IL-8 from individual rabbits infected with the WT strain or from uninfected rabbits. Colored dots correspond to micrographs with similar colored borders. Mean values are indicated with bars. (Micrographs) Ascomycin (FK520) Immunofluorescence micrographs of colonic sections from uninfected animals or infant rabbits infected with WT strain. Sections Ascomycin (FK520) were stained with a RNAscope probe to rabbit IL-8 (red) and an antibody to (green) and with DAPI (blue). Download FIG?S3, TIF file, 2.8 MB. Copyright ? 2020 Kuehl et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S1. Transposon library in 2a strain 2457T. Download Table?S1, XLSX file, 0.01 MB. Copyright ? 2020 Kuehl et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International Ascomycin (FK520) license. FIG?S4. Localization of mutants in infected infant rabbits. (A to C) Immunofluorescence micrographs of mutants in colonic tissue of infected rabbits 36 hpi. (A) Inset and white arrows show individual cells closely associated with the colonic epithelium. Blue, DAPI; green, FITC-conjugated anti-antibody; red, phalloidin-Alexa Fluor 568. Scale bars, 500 m (A to C). Download FIG?S4, EPS file, 2.8 MB. Copyright ? 2020 Kuehl et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S2. Strains, plasmids, and oligonucleotides. Download Table?S2, XLSX file, 0.01 MB. Copyright ? 2020 Kuehl et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. ABSTRACT species cause diarrheal disease globally. Shigellosis is typically characterized by bloody stools and colitis with mucosal damage and is the leading bacterial cause of diarrheal death worldwide. After the pathogen is orally ingested, it invades and replicates within the colonic epithelium through mechanisms that rely on its type III secretion system (T3SS). Currently, oral infection-based small animal models to study the pathogenesis of shigellosis are lacking. Here, we found that orogastric inoculation of infant rabbits with led to diarrhea and colonic pathology resembling that within human shigellosis. Fasting animals to inoculation improved the frequency of disease prior. The pathogen colonized the digestive tract, where both intraepithelial and luminal foci had been noticed. The intraepithelial foci most likely arise through Ascomycin (FK520) growing from cell to cell. Robust intestinal colonization, invasion from the colonic epithelium, and epithelial sloughing all needed the T3SS aswell as IcsA, one factor necessary for bacterial adhesion Ascomycin (FK520) and growing mRNA labeling, was higher in pets contaminated with wild-type versus mutant strains lacking in or T3SS, recommending that epithelial invasion promotes manifestation of the chemokine. Collectively, our results suggest that dental infection of baby rabbits offers a good experimental model for research from the pathogenesis.