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Methods Mol Biol. 2017;1493:163-170.

Using Heterologous COS-7 Cells to Identify Semaphorin-Signaling Components.

Author information

1
Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, 30 Convent Drive, Room 320, Bethesda, MD, 20892, USA.
2
Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, 30 Convent Drive, Room 320, Bethesda, MD, 20892, USA. sgutkind@ucsd.edu.
3
University of California San Diego and Moores Cancer Center, 3855 Health Sciences Drive, #0803, La Jolla, CA, 92093, USA. sgutkind@ucsd.edu.

Abstract

Semaphorins are a family of membrane-bound and secreted type of proteins which were initially identified as chemorepulsive axon guidance molecules. Plexins and neuropilins are two major receptor families of semaphorins, and their common downstream targets are the actin cytoskeleton and cell-to-extracellular matrix adhesions. Semaphorins promote the collapse of growth cones by inducing rapid changes in the cytoskeleton and disassembly of focal adhesion structures. When transfected with appropriate receptors, non-neuronal COS-7 cells exhibit a similar cell collapse phenotype upon semaphorin stimulation. This heterologous system using COS-7 cells has been developed and widely used to investigate semaphorin-signaling pathways. In this chapter, we describe a COS-7 collapse assay protocol used to identify semaphorin-signaling components and a method to produce recombinant class 3 semaphorin proteins.

KEYWORDS:

Actin cytoskeleton; COS-7 cells; Cell collapse; Neuropilin; Plexin; Semaphorin

PMID:
27787849
DOI:
10.1007/978-1-4939-6448-2_11
[Indexed for MEDLINE]

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