Abstract
Tumor cell motility and invasion rely on actin cytoskeleton rearrangements mediated by the activation of RhoGTPase signaling pathways. Invadopodia are membrane-degrading protrusions that mediate extracellular matrix degradation. Here, we provide procedures for imaging RhoGTPase biosensors in tumor cells during the formation of invadopodia and matrix degradation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bravo-Cordero JJ et al (2011) A novel spatiotemporal RhoC activation pathway locally regulates cofilin activity at invadopodia. Curr Biol 21:635–644
Bravo-Cordero JJ, Hodgson L, Condeelis J (2012) Directed cell invasion and migration during metastasis. Curr Opin Cell Biol 24:277–283
Oser M et al (2009) Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. J Cell Biol 186:571–587
Oser M et al (2010) Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia. J Cell Sci 123:3662–3673
Pertz O, Hodgson L, Klemke RL, Hahn KM (2006) Spatiotemporal dynamics of RhoA activity in migrating cells. Nature 440:1069–1072
Segall JE et al (1996) EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism. Clin Exp Metastasis 14:61–72
Kaptein LC, Greijer AE, Valerio D, van Beusechem VW (1997) Optimized conditions for the production of recombinant amphotropic retroviral vector preparations. Gene Ther 4:172–176
Hodgson L, Shen F, Hahn K (2010) Biosensors for characterizing the dynamics of rho family GTPases in living cells. Curr Protoc Cell Biol Chapter 14(Unit 14.11):11–26, Editorial board, Juan S. Bonifacino … [et al.]
Spiering D, Hodgson L (2012) Multiplex imaging of Rho family GTPase activities in living cells. Methods Mol Biol 827:215–234
Acknowledgments
We thank the Condeelis, Cox, Hodgson, and Segall laboratories for helpful discussions. This work was funded by GM093121 (J.J.B-C, Y.M., L.H.), T32 GM007491 (Y.M.), and CA150344 (J.C, J.J.B-C).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bravo-Cordero, J.J., Moshfegh, Y., Condeelis, J., Hodgson, L. (2013). Live Cell Imaging of RhoGTPase Biosensors in Tumor Cells. In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_22
Download citation
DOI: https://doi.org/10.1007/978-1-62703-538-5_22
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-537-8
Online ISBN: 978-1-62703-538-5
eBook Packages: Springer Protocols