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. 2017 Jan 13;8:13998. doi: 10.1038/ncomms13998

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Copyright © 2017, The Author(s)

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(a–e) Confocal microscopy analysis of Crb3 (a), aPKC (b), Par3 (c), villin (d) and actin (e) on the apical or lateral sides in control (Caco2 shNT, left panels) and EpCAM-depleted (Caco2 shEpCAM, right panels) cells. xy and xz views are presented showing the relocated apical markers at tricellular junctions. Scale bars, 5 μm. (f) TEM ultrastructural analysis of tricellular contacts in control (Caco2 shNT, left) and EpCAM-depleted (Caco2 shEpCAM, right) cells, showing microvilli at tricellular contacts. Arrowheads point to TJs. Longitudinal (xy) and transversal views (xz) are presented. BC, bicellular contact, N, nucleus. Scale bars xy, 1 μm and xz, 2 μm. (g–k) Confocal microscopy analysis of double immunostainings for Crb3 (green) and occluding (red) (g), Crb3 (green) and E-cadherin (red) (h), villin (green) and occluding (red) (i), villin (green) and E-cadherin (red) (j), and E-cadherin (green) and occluding (red) (k) on the apical or lateral sides in EpCAM-depleted (Caco2 shEpCAM) cells. xy and xz views are presented. Scale bars, 5 μm; high magnifications, 2 μm. Nuclei were detected with Hoechst 33342 staining (blue).