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

CIL:13719*  Cite 

PINK1-YFP R98F (green) is localized to mitochondria in the absence of the mitochondrial depolarizing agent CCCP (carbonyl cyanide-m-chlorophenyl hydrazone). The R98F mutation in PINK1-YFP partially inhibits proteolytic cleavage. When fixed cells are permeabilized with 0.005% digitonin, the antibody to GFP (red) (used to identify PINK1-YFP R98F) is unable to reach and bind PINK1-YFP R98F, but the antibody to Tom20 (white), a mitochondrial outer membrane marker, is able to bind its epitope. This demonstrates that PINK1 R98F is located inside mitochondria. PINK1-YFP R98F transfected HeLa cells were treated with DMSO 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, 4th row from top, and is part of a differential permeabilization assay to determine submitochondrial localization of PINK1 R98F 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
cervical carcinoma
Cellular Component
mitochondrial outer membrane translocase complex
mitochondrial inner membrane
mitochondrial outer membrane
Seok Min Jin
Michael Lazarou
Chunxin Wang
Lesley A. Kane
Derek P. Narendra
Richard J. Youle
J Cell Biol, 191: 933-942, 2010.
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
Processing History
brightness and contrast adjusted
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Spatial Axis Image Size Pixel Size
X 512px 0.186µm
Y 512px 0.186µm