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), IL22RA1 (rat, 1:50, FAB42941P; R&D Systems), OLFM4 (rabbit, 1:250, 39141; Cell Signaling Technology).

Techniques: Expressing, Isolation, Sequencing, Immunohistochemistry, Staining

Journal: Atherosclerosis

Article Title: IL-22 affects smooth muscle cell phenotype and plaque formation in apolipoprotein E knockout mice.

doi: 10.1016/j.atherosclerosis.2015.08.006

Figure Lengend Snippet: Fig. 1. Expression of the IL-22 receptor in the vascular wall. Sections from the aortic root of 12 week old (A) and 24 week old (B) Apoe/ mice were stained for the IL-22RA1 subunit of the IL-22 receptor and a-actin. Positive staining is shown in brown (DAB). (C) Carotid arteries from wild type C57Bl6 mice, with our without vascular injury were stained for IL- 22RA1 or myosin heavy chain (MyHC11) using immunofluorescence. IL-22RA1 and MyHC11 are shown in red, DAPI in blue, and elastic membranes in green autofluoresence. Exclusion of primary antibody served as negative control. Scale bars represent 100 mm.

Article Snippet: For analysis of aortic root plaques, frozen tissue sections were cut from the aortic root, frozen and stained using the following primary antibodies: rat anti-mouse IL-22RA1 antibody (R&D systems, Abingdon, UK), rabbit anti-mouse SMC a-actin antibody (Abcam, Cambridge, UK), rat anti-mouse MOMA-2 antibody (BMA Biomedicals, Augst, Switzerland), rabbit-anti-mouse Caldesmon (Abcam), rabbit-anti-mouse CD68 (Abcam).

Techniques: Expressing, Staining, Negative Control