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

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.

• 2021

• Jin, Seok Min
• Kane, Lesley A.
• Lazarou, Michael
• Narendra, Derek P.
• Wang, Chunxin
• Youle, Richard J.

Preparation: formaldehyde fixed tissue; detergent permeabilized
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: single-spot confocal microscopy
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein
Visualization methods: EYFP; primary antibody plus labeled secondary antibody
Processing history: brightness and contrast adjusted
Data qualification: Raw;spatialmeasurements;intensitiesquantitation

• Cell Image Library Group ID: 7051

• Homo sapiens

• Endothelial cell
• Mitochondrial inner membrane
• Mitochondrial outer membrane
• Mitochondrial outer membrane translocase complex
• Mitochondrion

• ATP binding
• C3HC4-type RING finger domain binding
• Calcium-dependent protein kinase activity
• Cervical carcinoma
• HeLa
• Intracellular protein kinase cascade
• Magnesium ion binding
• P-P-bond-hydrolysis-driven protein transmembrane transporter activity
• Protein phosphorylation
• Protein serine/threonine kinase activity
• Protein targeting to mitochondrion
• Response to stress
• Ubiquitin protein ligase binding

• data
• image

• No linguistic content; Not applicable

Samplenumber: 13719

Doi: https://doi.org/10.6075/J0J9654N

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL13719

Reference

• J Cell Biol, 191: 933-942, 2010. https://www.ncbi.nlm.nih.gov/pubmed/?term=21115803

Other resource

• BioStudies (previously JCB DataViewer): https://www.ebi.ac.uk/biostudies/studies/

Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International Public License

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2022-08-12