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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 unpermeabilized (not treated with detergent), antibodies to GFP (red) (used to identify YFP-WT PINK1), and Tom20 (white), a mitochondrial outer membrane marker, are unable to reach and bind their epitopes. YFP-WT PINK1 transfected HeLa cells were treated with CCCP (10µM) 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, top row, and is a control experiment for 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:13717, Homo sapiens, endothelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0XD10DQ
Type
image
Identifier
ark:/20775/bb1235301v
Language
No linguistic content
Subject
Intracellular protein kinase cascade HeLa Protein targeting to mitochondrion Cervical carcinoma Response to stress Magnesium ion binding P-P-bond-hydrolysis-driven protein transmembrane transporter activity Protein phosphorylation Oxidative phosphorylation uncoupler activity ATP binding Calcium-dependent protein kinase activity C3HC4-type RING finger domain binding Protein serine/threonine kinase activity Ubiquitin protein ligase binding Mitochondrial outer membrane Endothelial cell Mitochondrial outer membrane translocase complex Mitochondrion Mitochondrial inner membrane Homo sapiens Cell Image Library Group ID: 7051
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