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Integrin Dependent RhoB Activation Assay Using Leukocytes
Department of Experimental Medical Science, Lund University, Lund, Sweden.
Department of Experimental Medical Science, Lund University, Lund, Sweden.
Department of Experimental Medical Science, Lund University, Lund, Sweden.
Örebro University, School of Medical Sciences. Department of Experimental Medical Science, Lund University, Lund, Sweden.
2018 (English)In: Bio-protocol, E-ISSN 2331-8325, Vol. 8, no 23, article id e3105Article in journal (Refereed) Published
Abstract [en]

To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including RhoB. Here we describe an activation assay of RhoB in leukocytes migrating on ICAM-1Fc coated dishes using commercially available Rhoteikin coated agarose beads. Although this is a specific protocol for LFA-1 induced RhoB activation, it can also be used for RhoB activation induced by other soluble and insoluble factors.

Place, publisher, year, edition, pages
Bio-Protocol , 2018. Vol. 8, no 23, article id e3105
Keywords [en]
Leukocytes, LFA-1, RhoB, Small GTPases, Rhoteikin, Pull-down
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:oru:diva-72536DOI: 10.21769/BioProtoc.3105ISI: 000458028500010OAI: oai:DiVA.org:oru-72536DiVA, id: diva2:1289845
Funder
Swedish Research Council, K2010-80P-21592-01-4 K2010-80X-215917-01-4The Royal Swedish Academy of SciencesÅke Wiberg Foundation
Note

Funding Agencies:

Foundation Olle Engvist Byggmästare  

I & A Lundberg Research Foundation  

Jeanssons Foundation  

Kocks Foundation  

P & U Schybergs Foundation  

Gyllenstiernska Krapperup Foundation  

Gustav V 80 Jubilee Fund  

Österlund Foundation  

Nanna Svartz, Crafoord Awards Anna-Greta Crafoord Foundation  

Royal Physiographic Society of Lund 

Available from: 2019-02-19 Created: 2019-02-19 Last updated: 2019-02-19Bibliographically approved

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Svensson, Lena

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