Detail of Fig1C_aTubulin-Afadin
Project
Title
SIM images of tubulin immunofluorescence in Eph4 cells
Description
NA
Release, Updated
2018-11-14
License
CC BY
Kind
Image data
based on Experiment
File Formats
Data size
1.2 MB
Datatype
Microtubule immunofluorescence
Molecular Function (MF)
Biological Process (BP)
-
Cellular Component (CC)
cell cell junction
(
GO:0005911
)
Biological Imaging Method
XYZ Scale
XY: 0.04 micrometer/pixel, Z: 0.3 micrometer/slice
T scale
-
Image Acquisition
Experiment type
Immunofluorescence
Microscope type
FluorescenceMicroscope
Acquisition mode
Other
Contrast method
Fluorescence
Microscope model
Zeiss Elyra S.1 SIM (ELYRA S.1)
Detector model
-
Objective model
Plan Apochromat 100×, 1.46 NA oil im- mersion lens, 63×, 1.4 NA oil immersion lens, and 40×, 1.4 NA oil immer- sion lens
Filter set
-
Summary of Methods
See details in Yano et al. (2013) J Cell Biol, 203(4): 605-614.
Related paper(s)
Tomoki Yano, Takeshi Matsui, Atsushi Tamura, Masami Uji, Sachiko Tsukita (2013) The association of microtubules with tight junctions is promoted by cingulin phosphorylation by AMPK., The Journal of cell biology, Volume 203, Number 4, pp. 605-14
Published in 2013 Nov 25
- doi: 10.1083/jcb.201304194
-
pmid: 24385485
(Abstract) Epithelial cells characteristically have noncentrosomal microtubules that are arranged in the apicobasal direction. In this paper, we examined cell sheets formed by an epithelial (Eph4) cell line by structure illumination microscopy and found a previously not clearly described planar apical network of noncentrosomal microtubules (MTs) in which the sides of the MT bundles were associated with tight junctions (TJs). In a gel overlay assay with taxol-stabilized MTs, cingulin showed strong binding to MTs, and a domain analysis showed that this binding occurred through cingulin's N-terminal region. The association of planar apical MTs with TJs was compromised by cingulin knockdown (KD) or the expression of dephosphomimetic mutants of cingulin at its adenosine monophosphate-activated protein kinase (AMPK) target sites, whereas phosphorylation at these sites facilitated cingulin-tubulin binding. In addition, although wild-type colonies formed spheres in 3D culture, the cingulin KD cells had anisotropic shapes. These findings collectively suggest that the regulated cingulin-MT association has a specific role in TJ-related epithelial morphogenesis that is sensitive to metabolic homeostasis-related AMPK activity.(MeSH Terms)
- AMP-Activated Protein Kinases/metabolism[major]
- Amino Acid Sequence
- Animals
- Calcium/metabolism
- Cell Polarity
- Centrosome/metabolism
- Epithelial Cells/cytology
- Epithelial Cells/metabolism
- Epithelium/growth & development
- Epithelium/metabolism
- Fluorescence Resonance Energy Transfer
- Fluorescent Antibody Technique
- Gene Knockdown Techniques
- HEK293 Cells
- Humans
- Intercellular Junctions/metabolism
- Membrane Proteins/chemistry
- Membrane Proteins/metabolism[major]
- Mice
- Microtubules/metabolism[major]
- Models, Biological
- Molecular Sequence Data
- Morphogenesis
- Phosphorylation
- Phosphoserine/metabolism
- Protein Structure, Tertiary
- Sus scrofa
- Tight Junctions/metabolism[major]
- Tubulin/metabolism
Contact
Sachiko Tsukita
, Osaka University
, Graduate School of Frontier Biosciences and Medicine
, Laboratory of Biological Science
Contributors
Tomoki Yano, Takeshi Matsui, Atsushi Tamura, Masami Uji, Sachiko Tsukita
OMERO Dataset
OMERO Project
Source