SSBD:database

389-Watabe-PGE2discharge/Fig2A_10nM thumbnail

389-Watabe-PGE2discharge/Fig2A_10nM

Author reviewed
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 10 nM PGE2
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Project
389-Watabe-PGE2discharge
Dataset ID
Fig2A_10nM
Kind
Image data
Title
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 10 nM PGE2
Description
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells (COX-1 and COX-2 double knockout HeLa cells) treated with 10 nM PGE2 (prostaglandin E2) for 20 to 30 min. The data of 3 independent experiments are reposited.
Folder Name
Fig2A_10nM
File Format
.nd2
File Size
7MB
File Separation Method
3 independent images
Figure Number
Fig2A
Dimensions
576x576x1x3x1
X Scale
2.7 micrometer
Y Scale
2.7 micrometer
Z Scale
-
T Scale
-
Channel
3 channel
Quantitative Data
-
Paper(s)

Biosample

Description
Reporter cells; COX-1 and COX-2 (COX-DKO)-deficient HeLa cells which express Gs-coupled EP2 receptor and Booster-PKA
Organism
Homo sapiens (NCBITaxon:9606)
Strain
-
Cell
-
Cell line
HeLa cell (CLO:0003684)
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)
Intrinsic Variable
In the reporter cells, cyclooxygenases (COX1 and COX2) were knocked out by CRISPR/Cas9 to eliminate PGE2 synthesis.
Extrinsic Variable
EP2 receptor and Booster-PKA were expressed. 10 nM PGE2 application

Ontology

Anatomical Entity
-
UBERON
-
Biological Process
cell communication (GO:0007154)
signal transduction (GO:0007165)
Cellular Component
cytoplasm (GO:0005737)
Molecular Function
-
MeSH Anatomy
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)
MeSH BP
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)
MeSH CC
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)
MeSH MF
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)

Genetics

Gene
-
Protein / Probe
-Booster-PKA
Method / Genotype
transposition
Gene
PTGS1, PTGS2
Protein / Probe
-
Method / Genotype
gene knock out
Gene
-
Protein / Probe
Prostaglandin E2 receptor EP2 subtype
Method / Genotype
transposition
Gene
-
Protein / Probe
-OptoSTIM1 (CRY11clust)
Method / Genotype
transposition

Treatments

Reagent / Compound
prostaglandin E2
Concentration
10 nanomolar
Fold Dilution
-

Imaging Method

Imaging Method
fluorescence microscopy (FBbi:00000246)
FRET (FBbi:00000367)
Detection Method
wide-field detection
Visualization Method
Booster-PKA, a PKA biosensor, mKate2
Illumination Method
widefield illumination
Source of Contrast
-
Imaged Parameter
fluorescence emission
Sample Preparation Method
living tissue
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Cell Communication (D002450)
Cytoplasm (D003593)
Fluorescence Resonance Energy Transfer (D031541)
Signal Transduction (D015398)

Instrument

Body
Nikon ECLIPSE Ti2
Module
-
Light Source
Excelitas Technologies Corp. X-Cite TURBO LED light source+O4:AN4
Detector
Hamamartsu ORCA Fusion Digital CMOS camera
Objective
Nikon CFI Plan 4X
Filter Set
A 555BP10 excitation filter (Omega Optical, Brattleboro, VT, USA), an FF562Di03 dichroic mirror (Semrock, Rochester, NY, USA), and XF3024 (590DF35) (Omega Optical) and BLP01-633R-25 (Semrock) emission filters for mKOκ and mKate2, respectively.
Dichroic
see Filter Set