Detail of Figure4D_Brafv600Ecell_GAS_20210821_WT3



Project
Title
Live imaging of ISP-GAS WT BrafV600E melanoma cells under two-photon microscopy at 7 days after tumor implantation.
Description
Live imaging of ISP-GAS WT BrafV600E melanoma cells under two-photon microscopy at 7 days after tumor implantation.
Release, Updated
2022-11-23
License
CC BY
Kind
Image data
File Formats
.oib
Data size
100.6 MB

Organism
Mus musculus ( NCBI:txid10090 )
Strain(s)
C57BL/6 (mouse)
Cell Line
BrafV600E cell ( NA )

Datatype
-
Molecular Function (MF)
IFN binding ( GO:0019961 ) interferon-gamma binding ( GO:0019964 )
Biological Process (BP)
Cellular Component (CC)
Biological Imaging Method
fluorescence microscopy ( Fbbi:00000246 )
two-photon excitation microscopy
X scale
0.414 micrometer/pixel
Y scale
0.414 micrometer/pixel
Z scale
10 micrometer/slice
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 Tanaka T, et. al. (2021) Cell Struct Funct, Dec 22;46(2):103-111.
Related paper(s)

Taisei Tanaka, Yoshinobu Konishi, Hiroshi Ichise, Shinya Tsukiji, Michiyuki Matsuda, Kenta Terai (2021) A Dual Promoter System to Monitor IFN-gamma Signaling in vivo at Single-cell Resolution., Cell structure and function, Volume 46, Number 2, pp. 103-111

Published in 2021 Dec 22 (Electronic publication in Nov. 6, 2021, midnight )

(Abstract) IFN-gamma secreted from immune cells exerts pleiotropic effects on tumor cells, including induction of immune checkpoint and antigen presentation, growth inhibition, and apoptosis induction. We combined a dual promoter system with an IFN-gamma signaling responsive promoter to generate a reporter named the interferon sensing probe (ISP), which quantitates the response to IFN-gamma by means of fluorescence and bioluminescence. The integration site effect of the transgene is compensated for by the PGK promoter-driven expression of a fluorescent protein. Among five potential IFN-gamma-responsive elements, we found that the interferon gamma-activated sequence (GAS) exhibited the best performance. When ISP-GAS was introduced into four cell lines and subjected to IFN-gamma stimulation, dose-dependency was observed with an EC50 ranging from 0.2 to 0.9 ng/mL, indicating that ISP-GAS can be generally used as a sensitive biosensor of IFN-gamma response. In a syngeneic transplantation model, the ISP-GAS-expressing cancer cells exhibited bioluminescence and fluorescence signals in an IFN-gamma receptor-dependent manner. Thus, ISP-GAS could be used to quantitatively monitor the IFN-gamma response both in vitro and in vivo.Key words: in vivo imaging, tumor microenvironment, interferon-gamma, dual promoter system.
(MeSH Terms)

Contact
Kenta Terai , kyoto University , Graduate School of Medicine , Department of Pathology and Biology of Diseases
Contributors
Kenta Terai, Taisei Tanaka

OMERO Dataset
OMERO Project
Source