Journal: Cellular and Molecular Gastroenterology and Hepatology
Article Title: Human AGR2 Deficiency Causes Mucus Barrier Dysfunction and Infantile Inflammatory Bowel Disease
doi: 10.1016/j.jcmgh.2021.07.001
Figure Lengend Snippet: Depletion of gel-forming mucins MUC2, MUC5AC, and MUC6 in gastrointestinal tissues of the patients. Fluorescent immunohistochemistry staining of formalin-fixed paraffin-embedded sections showing depletion of MUC2 (green, clone F-2) in small and large bowel, as well as MUC5AC (green, clone CLH2) and MUC6 (red, clone CLH5) in the stomach. Nuclei were stained with Hoechst (blue). Patient 2 shows a small group of cells positive for MUC6 ( arrowhead ). Representative images are shown for patients 1 and 2, 1 of 3 noninflamed controls, and 1 of 5 inflamed controls for small bowel. For large bowel, representative images are shown for 1 of 3 noninflamed controls and 1 of 6 inflamed controls. Representative, high-magnification (63× objective) images of small-bowel crypts show the reduced size of goblet cell thecae in the patients with staining for TFF3 (red, clone B-1) and MUC2 (green, clone F-2). All other images were acquired with a 10× objective. For gastric sections, MUC5AC staining was replicated across 3 noninflamed controls and 4 inflamed controls, while MUC6 staining was replicated across 2 noninflamed controls and 2 inflamed controls. Identical thresholding parameters were applied to the images from each respective tissue. Inflamed conditions shown in the figure are celiac for small bowel, appendicitis for large bowel, and H pylori for gastric. Staining for every marker was replicated a minimum of 3 independent times. Scale bars : 100 μm (low magnification); 10 μm (high magnification).
Article Snippet: Anti-mucin 6/MUC6 antibody (CLH5) Alexa Fluor 546 , Santa Cruz (Dallas, TX) , sc-33668 AF546 , IHC.
Techniques: Immunohistochemistry, Staining, Formalin-fixed Paraffin-Embedded, Marker