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. 2018 Aug 14;6(4):477–493.e1. doi: 10.1016/j.jcmgh.2018.08.001

Figure 2.

Figure 2

Brush border and trafficking defects in Munc18-2 KO organoids. (A) Detection of alkaline phosphatase (AP) enzymatic activity (top), Cd10 (middle), and Dpp4 immunostaining (bottom) on paraffin sections of WT and Munc18-2 KO organoids. Overview (left) and magnified villus regions (right) are shown. White arrows show subapical intracellular accumulation of brush-border components. (B) TEM analysis of the apical region. Crypt regions (top) show accumulation of empty vesicles in the Munc18-2 KO. Villus regions (bottom) show the presence of tubulovesicular structures and (auto)lysosomes (arrow). No MVIs are found under expansion conditions. (C–E) Quantification of microvilli (MV) length, width, and density. In total, 70 MVs were measured in n = 7 organoids. Mean values ± SD are shown, P values were determined by t test. (F) Paneth cells (top) and goblet cells (bottom) show some immature aspects in Munc18-2 KO organoids, with reduced electron density of Paneth cell granules and decreased mucus content in goblet cells. Scalebars: (A) 10 μm, (B and F) 2 μm. All staining was confirmed in at least 2 independent experiments.