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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 0.25% Triton X-100 (TX-100), 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 0.25% Triton X-100. 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, 3rd 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. 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:13722, Homo sapiens, endothelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J09S1PRP
Type
image
Identifier
ark:/20775/bb1610733p
Language
No linguistic content
Subject
Intracellular protein kinase cascade Calcium-dependent protein kinase activity Response to stress Protein targeting to mitochondrion HeLa Ubiquitin protein ligase binding ATP binding C3HC4-type RING finger domain binding Magnesium ion binding Cervical carcinoma Oxidative phosphorylation uncoupler activity Protein serine/threonine kinase activity P-P-bond-hydrolysis-driven protein transmembrane transporter activity Protein phosphorylation Mitochondrial outer membrane translocase complex Endothelial cell Mitochondrion Mitochondrial inner membrane Mitochondrial outer membrane Homo sapiens Cell Image Library Group ID: 7051
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