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

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© 2018 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Confocal analysis of brush border and trafficking defects in Munc18-2 KO organoids. (A) Defective deposition of brush-border F-actin as shown by phalloidin staining (red). Zonula occludens-1 (ZO-1) co-staining (green) shows that tight junction–associated F-actin is not affected. Single confocal sections (with DAPI-stained nuclei in white). The magnified z-projected images (bottom) show a planar view onto the brush border. WT organoids show a continuous brush border (asterisks) that is interrupted in the Munc18-2 KO (white arrowheads). (B) Confocal analysis of F-actin (red) and Stx3 (green). Stx3 shows a diffuse subapical localization in the Munc18-2 KO (white arrow). Bottom: Magnified z-projected images. Scale bars: 10 μm. Staining was confirmed in 3 independent experiments.