Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

PMC Copyright notice

(a) Statistical analysis of cyst morphology in Caco2 shNT (white), Caco2 shEpCAM#1 (grey) and Caco2 shEpCAM#2 (black) cells. Three independent experiments were performed. N_(Caco2shNT)=779 cysts, N_{(Caco2shEpCAM#1)}=1,049 cysts, N_{(Caco2shEpCAM#2)}=1,686 cysts. Unpaired t-tests, ****P<0.0001, ***P<0.001, **P<0.01, *P<0.1. Percentage of Caco2shNT cells with multilumen cysts=4.5±2.5%, no lumen cysts=0.13±0.2%, multilayered cysts=0.9±1.7%, unshaped cysts=4.1±3.0% and reversed polarity cysts=0.2±0.2%. Percentage of Caco2shEpCAM#1 cells with multilumen cysts=28.6±4.9%, no lumen cysts=0.2±0.3%, multilayered cysts=10.9±3.0%, unshaped cysts=16.7±4.5% and reversed polarity cysts=9.6±2.2%. Percentage of Caco2shEpCAM#2 cells with multilumen cysts=26.6±8.5%, no lumen=3.0±1.1%, multilayered cysts=18.5±6.3%, unshaped cysts=15.8±1.5% and reversed polarity cysts=16.1±4.7%. Values are mean±s.d. (b–e) Confocal analysis of E-cadherin (b), actin (c), ezrin (d) and Crb3 (e) in 21 days 3D Matrigel cultures of control (Caco2 shNT) and EpCAM-silenced (Caco2 shEpCAM) cells. (b(c)) Arrowheads point toward weakened E-cadherin-based cell adhesions. (b(d)) Arrowheads point toward basal redirection of lateral E-cadherin. (d–e) Arrowheads point toward apical marker mislocalization on the outer side of the cysts, reflecting the loss of apico-basal polarity in shEpCAM cells. Nuclei were detected with Hoechst 33342 staining (blue). Scale bars, 10 μm. (f,g) Confocal analysis of double immunostainings for villin (green) and ZO-1 (red) (f), or E-cadherin (green) and ezrin (red) (g) in 21 days 3D Matrigel cultures of control (Caco2 shNT) and EpCAM-silenced (Caco2 shEpCAM) cells. Higher magnifications on EADs are presented in the lower panels. Nuclei were detected with Hoechst 33342 staining (blue). Scale bars, 20 μm.