366-Ebata-TERTlocalize/Fig2_FigS2_TERT_2
Author reviewedTime-lapse images of HeLa cells with or without expressing the TERT constructs during apoptosis
Project
Dataset ID
Fig2_FigS2_TERT_2
Kind
Image data
Title
Time-lapse images of HeLa cells with or without expressing the TERT constructs during apoptosis
Description
Time-lapse images of HeLa cells with or without expressing the TERT (Telomerase reverse transcriptase) constructs during apoptosis induced by oxidative stress. This dataset is composed of 16 series of dataset; series1-series16, each of which is 3-day long time-lapse image of 7 x 7 tiling data. Channel2; bright field, Channel4; SYTOX Orange, Channel5; merge. The 16 series consist of 4 genotypes and 4 experimental conditions.
Genotypes:
Series1-Series4; without transfection,
Series5-Series8; TERT transfection,
Series9-Series12; mutant TERT (R3ER6E) transfection,
Series13-Series16; mutant TERT (Y707F) transfection.
Experimental conditions:
Series1, 5, 9, 13; SYTOX Orange was used as an apotosis marker,
Series2, 6, 10, 14; YO-PRO-1 was used as an apotosis marker,
Series3, 7, 11, 15; SYTOX Orange imaging with treatment of Z-VAD-fmk,
Series 4, 8, 12, 16; SYTOX Orange imaging with treatment of GSK'872.
Folder Name
Fig2_FigS2_TERT_2
File Format
.nd2
File Size
589GB
File Separation Method
day1, day2, day3
Figure Number
Fig2, FigS2
Dimensions
7107x7107x1x3x72
X Scale
0.324 micrometer
Y Scale
0.324 micrometer
Z Scale
-
T Scale
1 hour
Channel
3 channel
Quantitative Data
-
Paper(s)
Biosample
Description
Wild-type HeLa cells TERT (Telomerase reverse transcriptase)-transfected HeLa cells, TERT mutant-transfected HeLa cells (R3E/R6E mutation or Y707F mutation)
Organism
Homo sapiens (NCBITaxon:9606)
Strain
-
Cell
-
Cell line
HeLa cell (CLO:0003684)
MeSH
Intrinsic Variable
HeLa cells stabely expressing TERT (Telomerase reverse transcriptase) or mutant TERT (R3E/R6E or Y707F)
Extrinsic Variable
SYTOX Orange or YO-PRO-1 application, which stains dead cells. sodium percarbonate pretreatment as oxidative stress.
Ontology
Anatomical Entity
-
UBERON
-
Biological Process
regulation of apoptotic process (GO:0042981)
Cellular Component
mitochondrion (GO:0005739)
nucleus (GO:0005634)
Molecular Function
telomerase activity (GO:0003720)
MeSH Anatomy
MeSH BP
MeSH CC
MeSH MF
Genetics
Gene
TERT
Protein / Probe
Telomerase reverse transcriptase
Method / Genotype
transfection
Gene
-
Protein / Probe
-SYTOX Orange
Method / Genotype
-
Gene
-
Protein / Probe
-YO-PRO-1
Method / Genotype
-
Treatments
Reagent / Compound
Sodium percarbonate
Concentration
267 micromolar
Fold Dilution
-
Reagent / Compound
Z-VAD-fmk
Concentration
50 micromolar
Fold Dilution
-
Reagent / Compound
GSK'872
Concentration
5 micromolar
Fold Dilution
-
Imaging Method
Imaging Method
fluorescence microscopy (FBbi:00000246)
time lapse microscopy (FBbi:00000249)
Detection Method
wide-field detection
Visualization Method
SYTOX Orange, YO-PRO-1
Illumination Method
widefield illumination
Source of Contrast
-
Imaged Parameter
-
Sample Preparation Method
living tissue
MeSH
Instrument
Body
inverted microscope (Ti, Nikon)
Module
-
Light Source
X-Cite XLED1, Lumen Dynaics
Detector
ImagEM X2-1K EM-CCD, Hamamatsu
Objective
Plan Apo Lambda 40×/0.95, Nikon
Filter Set
[CFP-2432C (for SYTOX-Blue or Cellstain Hoechst 33342; Semrock, Rochester, NY, USA), GFP-3035D (for YO-PRO-1; Semrock), LF514-B (for mVenus; Semrock), TRITC-A-Basic (for SYTOX-Orange; Semrock), and Cy5-4040C (for MitoTracker Deep Red FM; Semrock)].
Dichroic
-