Mucus-expelling GCs exhibit TJ reorganization and delocalization of cell polarity markers and express luminally accessible Ecad. (A) Intestinal tissues were fixed and stained with WGA and for accessible (acc) hEcad and nuclei. Mucus-expelling and mucus-filled GCs expressing luminally accessible hEcad were quantified. Error bars indicate SD. n = 30 villi from three mice. (right) Stack projection of intestinal epithelium showing an accessible hEcad⁺ GC expelling its mucosal content (arrows; a) and an accessible hEcad⁻ GC full of mucus (b). White dashed lines outline the border of GCs. (B) Intestinal tissues were fixed and stained with WGA and for accessible hEcad and TJ proteins (red). (top) Accessible hEcad of a GC (arrows) colocalizes with thickened TJ protein occludin. (bottom) TJ protein ZO-1 is thickened on the apical side of a GC (arrows) with accessible hEcad. (C, top) PAR-3 is depolarized around mucus-expelling GCs (asterisks). (bottom) PKCζ is delocalized from the apical region on mucus-expelling GCs (asterisks). (B and C) Stack projections of whole mount tissues are shown. (D) Intestinal tissues were fixed and stained with WGA and for accessible hEcad, total hEcad, and nuclei. (top) A GC (asterisks) presents an enrichment of total hEcad along its lateral membranes. (bottom) XZ and YZ sections at selected position. Pictures are representative of three mice. Bars, 10 µm.