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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 unpermeabilized (not treated with detergent), antibodies to GFP (red) (used to identify PINK1-YFP R98F), and Tom20 (white), a mitochondrial outer membrane marker, are unable to reach and bind their epitopes. PINK1-YFP R98F transfected HeLa cells were treated with DMSO for 3 hrs, fixed with 4% paraformaldehyde and not permeabilized. 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:13718, Homo sapiens, endothelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0MS3RGG
Type
image
Identifier
ark:/20775/bb73104424
Language
No linguistic content
Subject
HeLa Protein phosphorylation ATP binding Protein targeting to mitochondrion Magnesium ion binding Calcium-dependent protein kinase activity P-P-bond-hydrolysis-driven protein transmembrane transporter activity C3HC4-type RING finger domain binding Protein serine/threonine kinase activity Cervical carcinoma Ubiquitin protein ligase binding Intracellular protein kinase cascade Response to stress Mitochondrial outer membrane Mitochondrion Endothelial cell Mitochondrial inner membrane Mitochondrial outer membrane translocase complex Homo sapiens Cell Image Library Group ID: 7051
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