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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:13720*  Cite 
Description

YFP-WT PINK1 (green) shows mitochondrial localization in HeLa cells following treatment with the mitochondrial depolarizing agent CCCP (carbonyl cyanide m-chlorophenyl hydrazone). When fixed cells are permeabilized with 0.005% digitonin, the antibody to GFP (red) (used to identify YFP-WT PINK1) and the antibody to Tom20 (white), a mitochondrial outer membrane marker, are able to bind their epitopes. This demonstrates that when mitochondrial membrane potential is dissipated, YFP-WT PINK1 accumulates on the outer mitochondrial membrane. YFP-WT PINK1 transfected HeLa cells were treated with CCCP (10µM) for 3 hrs, fixed with 4% paraformaldehyde and permeabilized with 0.005% digitonin. Primary antibodies used were: anti-GFP polyclonal Ab (Invitrogen) and anti-Tom20 mAb (BD); secondary antibodies used were: Alexa Fluor 594 and 647. Imaging was performed on an LSM510 Meta (Carl Zeiss, Inc) with a 63x 1.4 NA oil differential interference contrast Plan Apo objective. Image contrast and brightness were adjusted in the LSM image browser (Zeiss). This image corresponds to Fig 4c, 2nd row from top, and is part of a differential permeabilization assay to determine submitochondrial localization of PINK1 and that is further described in Fig 4c of J Cell Biol, 191: 933-942, 2010. Images in Fig 4 include CIL#s 13733, 13734, 13729, 13730, 13731, 13732, 13717, 13718, 13719, 13720, 13721, 13722, 13723, 13724.

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
endothelial cell
Cell Line
HeLa
cervical carcinoma
Cellular Component
mitochondrion
mitochondrial outer membrane translocase complex
mitochondrial inner membrane
mitochondrial outer membrane
Attribution
Names
Seok Min Jin
Michael Lazarou
Chunxin Wang
Lesley A. Kane
Derek P. Narendra
Richard J. Youle
Published
J Cell Biol, 191: 933-942, 2010.
Pubmed
21115803
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13720
Archival Resource Key (ARK)
ark:/b7295/w9cil13720
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EYFP
primary antibody plus labeled secondary antibody
Processing History
brightness and contrast adjusted
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 512px 0.186µm
Y 512px 0.186µm