Alternate header for print version

This image is copyright protected. Any public or private use of this image is subject to prevailing copyright laws. Please contact the content provider of this image for permission requests.
*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:16217*  Cite 

The unique-6 (U6) motif of the MAGUK protein, ZO-1, inhibits tight junction localization of GFP fusion proteins containing the SH3-GUK module. Stable MDCK II cell lines expressing GFP fusion proteins encoding GFP alone (A and B), ZO1 FL (full-length ZO-1; C and D), U5-GUK (E and F), SH3-U5-GUK (G and H), SH3-U5-GUK containing a GUK domain mutation that inhibits hairpin formation (I and J), U6 (K and L), U5-GUK-U6 (M and N), and SH3-U5-GUK-U6 (O and P) were plated on glass coverslips and grown to confluence. Cells were fixed and stained with an antisera specific to the endogenous ZO-1. The distribution of the GFP fusion (green) and ZO-1 (red) were observed by wide-field microscopy. Bar, 10 µm.

Technical Details

MDCK-II cells were plated on glass coverslips at a low density (1.0 x 10[4] cells/ml) and cultured for 2 d before processing or until cells had just reached confluence. Cells were fixed in 4% paraformaldehyde, permeabilized in 0.2% Triton X-100 and incubated in rat anti-ZO1 hybridoma supernatant (R40.76) followed by Cy3-conjugated donkey anti-rat secondary antibody. Cells were mounted in Mowiol with 1.0% n-propyl gallate. Wide-field images were acquired on a Nikon E800 microscope using 60x or 100x Plan Apo lenses and an Orca ER cooled CCD camera controlled with the Metamorph Imaging software package. Filter sets used include the Piston GFP Bandpass #41025 for acquisition of GFP images and the special green filter set #C4607 for Cy3 images. Figure 4 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
widefield illumination
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
Processing History
Data Qualifiers
processed data
Sample Preparation
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro