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Journal: Cellular and Molecular Gastroenterology and Hepatology
Article Title: IL22 Inhibits Epithelial Stem Cell Expansion in an Ileal Organoid Model
doi: 10.1016/j.jcmgh.2018.06.008
Figure Lengend Snippet: Il22ra1 is expressed heterogeneously throughout the crypt. ( A ) The Il22ra1 gene expression profile characterized in FACS-isolated total epithelium (CD326+), absorptive/goblet differentiated cells (Sox9-EGFP neg ), TA progenitor cells (Sox9-EGFP sublow ), ISCs (Sox9-EGFP low ), enteroendocrine/tuft cells (Sox9-EGFP high ), and Paneth cells (Sox9-EGFP high , CD24 high ). Technical replicate n = 3; biological N = 3 mice. Significance was calculated by 1-way analysis of variance with Tukey multiple comparisons; bars that are not connected by the same letter are statistically significant ( P < .05). ( B ) Left : t-SNE analysis of single-cell RNA sequence analysis of mouse small intestinal epithelium. Each color represents a different population defined in the original analysis based on lineage-specific transcriptomic signatures. Right : Table depicts the number of cells in each lineage category expressing IL22ra1. ( C ) Cells from the t-SNE profiles in the graph in panel B are highlighted specifically for the expression of IL22ra1 levels in all epithelial cells. Darker shades of grey represent higher expression levels. Pink circles represent no expression. ( D ) The same analysis in panel C except only ISCs are shown. ( E ) The same analysis in panel C except only TA progenitors are shown. ( F ) Representative immunohistochemistry of IL22RA1 (red) and cell nuclei (blue) in a mouse ileal crypt. ( G ) FACS analysis of fixed cell populations described in panel A stained for IL22RA1. Technical replicate n = 3; biological N = 3 mice. Bars represent parts of whole. EC, enterocyte; EE, enteroendocrine; EEC, enteroendocrine; Max, maximum; Min, minimum.
Article Snippet: Primary antibodies and dilutions used were as follows: KI67 (rabbit, 1:100, M7249; Dako), LYZ (goat, 1:500, sc-12091; Santa Cruz, Dallas, TX), CD326 epithelial cell adhesion molecule (EPCAM)/CD326 (rat, 1:500, H8201, clone G8.8; Biolegend, San Diego, CA),
Techniques: Expressing, Isolation, Sequencing, Immunohistochemistry, Staining