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

CIL:16220*  Cite 

ZO-1 polypeptides lacking the unique-6 (U6) motif are concentrated within disorganized strands on the lateral surface of MDCK cells. MDCK II cells transfected with deltaU6 (A–F) or ZNG (aa1-806) ( (G–H) were plated on filter inserts and grown in the presence (A–C) or absence (D–F and G–H) of doxycycline for 10 d. The ectopic strands were not present in uninduced cells (A-C). Cell were then fixed and stained with an antibody against c-myc (A, D, and G) and a sera specific for the endogenous ZO-1 (B, E, and H). The images shown are projections of 5.0 µm confocal stacks that have been rotated by 48°. 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 presence or absence of doxycyclin for 10 d before processing. Cells were fixed for 30 min in ethanol on ice, permeabilized in 0.2% Triton X-100 and incubated in anti-myc antibody and rat anti-ZO1 hybridoma supernatant (R40.76) followed by Cy3-conjugated donkey anti-rat secondary antibody and Cy2 or Cy5-conjugated secondary antibody specific for anti-myc. 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 7 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