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. 2018 Dec 5;8(23):e3105.
doi: 10.21769/BioProtoc.3105.

Integrin Dependent RhoB Activation Assay Using Leukocytes

Janne Lehtonen  1 , Katarzyna Potrzebowska  1 , Malin Samuelsson  1 , Lena Svensson  1   2
Affiliations

Integrin Dependent RhoB Activation Assay Using Leukocytes

Janne Lehtonen et al. Bio Protoc. .

Abstract

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.

Keywords: LFA-1; Leukocytes; Pull-down; RhoB; Rhoteikin; Small GTPases.

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Conflict of interest statement

Competing interestsThe authors declare no competing interest.

References

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    1. Samuelsson M., Potrzebowska K., Lehtonen J., Beech J. P., Skorova E., Uronen-Hansson H. and Svensson L.(2017). RhoB controls the Rab11-mediated recycling and surface reappearance of LFA-1 in migrating T lymphocytes. Sci Signal 10(509). - PubMed

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