Development of a Kinetic Assay for Late Endosome Movement

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14110%2F14%3A00077529" target="_blank" >RIV/00216224:14110/14:00077529 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1177/1087057114524278" target="_blank" >http://dx.doi.org/10.1177/1087057114524278</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1177/1087057114524278" target="_blank" >10.1177/1087057114524278</a>

Result language
angličtina
Original language name
Development of a Kinetic Assay for Late Endosome Movement
Original language description
Automated imaging screens are performed mostly on fixed and stained samples to simplify the workflow and increase throughput. Some processes, such as the movement of cells and organelles or measuring membrane integrity and potential, can be measured onlyin living cells. Developing such assays to screen large compound or RNAi collections is challenging in many respects. Here, we develop a live-cell high-content assay for tracking endocytic organelles in medium throughput. We evaluate the added value ofmeasuring kinetic parameters compared with measuring static parameters solely. We screened 2000 compounds in U-2 OS cells expressing Lamp1-GFP to label late endosomes. All hits have phenotypes in both static and kinetic parameters. However, we show thatthe kinetic parameters enable better discrimination of the mechanisms of action. Most of the compounds cause a decrease of motility of endosomes, but we identify several compounds that increase endosomal motility.
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CE - Biochemistry
OECD FORD branch
—

Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2014
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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