Detail of Figure3d_gB2-gB2



Project
Title
Time-lapse imaging of the FRET efficiency of γB2–γB2 by acceptor photobleaching in HEK293T cells.
Description
Time-lapse imaging of the FRET efficiency of γB2–γB2 by acceptor photobleaching in HEK293T cells.
Release, Updated
2022-11-23
License
CC-BY
Kind
Image data
File Formats
.oib
Data size
108.9 MB

Organism
Homo sapiens ( NCBITaxon:9606 )
Strain(s)
-
Cell Line
HEK293T ( CLO_0037271 )

Datatype
-
Molecular Function (MF)
Biological Process (BP)
homophilic cell adhesion via plasma membrane adhesion molecules ( GO:0007156 )
Cellular Component (CC)
Biological Imaging Method
time lapse microscopy ( Fbbi:00000249 )
X scale
0.138 micrometer/pixel
Y scale
0.138 micrometer/pixel
Z scale
-
T scale
-

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Kanadome T, et. al. (2021) Sci Rep, 2021 Nov 15;11(1):22237.
Related paper(s)

Takashi Kanadome, Natsumi Hoshino, Takeharu Nagai, Tomoki Matsuda, Takeshi Yagi (2021) Development of FRET-based indicators for visualizing homophilic trans interaction of a clustered protocadherin., Scientific reports, Volume 11, Number 1, pp. 22237

Published in 2021 Nov 15 (Electronic publication in Nov. 15, 2021, midnight )

(Abstract) Clustered protocadherins (Pcdhs), which are cell adhesion molecules, play a fundamental role in self-recognition and non-self-discrimination by conferring diversity on the cell surface. Although systematic cell-based aggregation assays provide information regarding the binding properties of Pcdhs, direct visualization of Pcdh trans interactions across cells remains challenging. Here, we present Forster resonance energy transfer (FRET)-based indicators for directly visualizing Pcdh trans interactions. We developed the indicators by individually inserting FRET donor and acceptor fluorescent proteins (FPs) into the ectodomain of Pcdh molecules. They enabled successful visualization of specific trans interactions of Pcdh and revealed that the Pcdh trans interaction is highly sensitive to changes in extracellular Ca(2+) levels. We expect that FRET-based indicators for visualizing Pcdh trans interactions will provide a new approach for investigating the roles of Pcdh in self-recognition and non-self-discrimination processes.
(MeSH Terms)

Contact
Tomoki Matsuda, Takeshi Yagi , Osaka University, Osaka University , Department of Biomolecular Science and Engineering , Graduate School of Frontier Biosciences , Laboratories for Integrated Biology,
Contributors

OMERO Dataset
OMERO Project
Source