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Description
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. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Jin, Seok Min; Lazarou, Michael; Wang, Chunxin; Kane, Lesley A.; Narendra, Derek P.; Youle, Richard J. (2021). CIL:13719, Homo sapiens, endothelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0J9654N
Type
image
Identifier
ark:/20775/bb3385491h
Language
No linguistic content
Subject
ATP binding Protein serine/threonine kinase activity Protein targeting to mitochondrion Calcium-dependent protein kinase activity P-P-bond-hydrolysis-driven protein transmembrane transporter activity Protein phosphorylation Intracellular protein kinase cascade Response to stress Magnesium ion binding Cervical carcinoma C3HC4-type RING finger domain binding Ubiquitin protein ligase binding HeLa Mitochondrial inner membrane Mitochondrion Mitochondrial outer membrane translocase complex Mitochondrial outer membrane Endothelial cell Homo sapiens Cell Image Library Group ID: 7051
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