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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:16222*  Cite 

Transmembrane proteins of the tight junction (occludin, claudin-2, JAM-A), but not cytosolic proteins (cingulin, ZO-2, alpha-catenin), are incorporated into the ectopic strands formed by delta-U6 constructs. MDCK cells expressing delta-U6 (Supplementary Figures) or myc-ZNG (aa 1-806) were plated at low density in DOX-free media, fixed after 4 d and stained with antibodies against c-myc (B, D, F, H, J, and L) and antisera recognizing the transmembrane proteins occludin (A), claudin-2 (C), and JAM-A (E) or the cytosolic proteins cingulin (G), ZO-2 (I), and alpha-catenin (K). The images shown are maximum projections of 2.5–5.0-µm confocal stacks. Bar, 10 µm.

Technical Details

MDCK-II tet off cells were plated on glass coverslips at a low density (1.0 x 10[4] cells/ml) and incubated in the absence of doxycyclin for 4 d before processing. Cells were fixed for 30 min in ethanol on ice, permeabilized in 0.2% Triton X-100 and incubated in mouse anti-myc 9E10 or rabbit anti-myc (MBL International;1:2,500), and mouse anti-ZO-2 3E8D9 (Zymed; 1:500), rabbit anti-cingulin (Sandra Citi, Geneva; 1:500), mouse anti-occludin (Zymed; 1:300), mouse anti-claudin-2 (Zymed; 1:200), rabbit anti-canine JAM-1 (Klaus Ebnet, Munster; 1:400) and α-catenin (Sigma-Aldrich; 1:2000) followed by Cy2, Cy3 or Cy5-conjugated secondary antibodies. Cells were mounted in Mowiol with 1.0% n-propyl gallate. Confocal images were acquired on a Zeiss LSM510 Meta using a 100x Plan Apo lens (Thornwood, NY). Confocal Stacks (E,F,G,H,M,N) and image projections were generated with Zeiss LSM Image Browser version 3.2. Contrast adjustment and montages were generated using Adobe Photoshop (version 6.0; San Jose, CA). Figure 9 in Mol Biol Cell (2006). 18:721-731.

Biological Sources
NCBI Organism Classification
Canis lupus familiaris
Cell Type
epithelial cell
Cell Line
Cellular Component
tight junction
Biological Context
Biological Process
tight junction assembly
Molecular Function
protein binding
Alan S. Fanning
Brent P. Little
Christopher Rahner
Darkhan Utepbergenov
Zenta Walther
James M. Anderson
Mol Biol Cell (2006). 18:721-731.
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
c-MYC peptide tag
Processing History
Data Qualifiers
processed data
Sample Preparation
ethanol fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro