The Epithelial-to Mesenchymal Transition, 1st ed. 2021
Methods and Protocols
Methods in Molecular Biology Series, Vol. 2179

This volume details a comprehensive range of methods for imaging  epithelial-to-mesechymal transition (EMT)/MET in in vivo systems, and methods to leverage these systems to dissect the underlying mechanisms. Chapters guide readers through studying different features of epithelial-mesenchymal plasticity, past and future research of the EMT, in vivo systems, and in vivo imaging. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The Epithelial-to Mesenchymal Transition: Methods and Protocols aims to provide methods in EMT will help to unite and drive research in this exciting field forwards.

Part I: Perspectives on the Past and Future of EMT Research

1. A Spectrum of Cell States during the Epithelial-To-Mesenchymal Transition

Erica J. Hutchins and Marianne E. Bronner

2. Perspective on Epithelial Mesenchymal Transitions in Embryos

3. EMT, One of Many Morphological Transitions in Cellular Phase Space

Denise J. Montell

4. Are you Interested or Afraid of Working on EMT?

5. Partial EMT/MET: An Army of One

Sofiane Hamidi, Hiroki Nagai, and Guojun Sheng

6. EMT; An Update

Jean Paul Thiery

Part II: Review on Mesenchymal-to-Epithelial Transitions

7. Mesenchymal to Epithelial Transitions in Development and Cancer

John-Poul Ng-Blichfeldtand Katja Röper

Part III: Methods to Study EMT in In Vivo Systems

8. Live Imaging of Epithelial-mesenchymal Transition in Mesoderm Cells of Gastrulating Drosophila Embryos

Lingkun Gu and Mo Weng

Zain Alhashem, Macarena Alvarez-Garcillan Portillo, Ravinand Htun, Anton Gauert, Luis Briones Montecinos, Steffen Härtel, and Claudia Linker

10. Live Imaging of the Neural Crest Cell Epithelial-to-Mesenchymal Transition in the Chick Embryo

Mary Cathleen McKinney and Paul M. Kulesa

11. Exploiting Drosophila melanogaster Wing Imaginal Disc Eversion to Screen for New EMT Effectors

Sofia Golenkina, Rosemary Manhire-Heath, and Michael J. Murray

12. Visualizing Mouse Embryo Gastrulation Epithelial-mesenchymal Transition through Single Cell Labeling Followed by ex vivo Whole Embryo Live Imaging

Wallis Nahaboo, Bechara Saykali, Navrita Mathiah, and Isabelle Migeotte

13. Fluorescence Recovery after Photobleaching to Study the Dynamics of Membrane Bound Proteins in vivo using the Drosophila Embryo

Joshua Greigand Natalia A. Bulgakova

14. Methods to Generate and Assay for Distinct Stages of Cancer Metastasis in adult Drosophila melanogaster  

Jamie Adams, Andreu Casali, and Kyra Campbell

Part IV: In Vivo Imaging of MET

15. 4D Live-Imaging and Analysis of Chick Embryo Somites

Gi Fay Mok, James McColl_, and Andrea Münsterberg

16. In vivo Analysis of the Mesenchymal-to-Epithelial Transition during Chick Secondary Neurulation

Elena Gonzalez-Gobartt, Guillaume Allio, Bertrand Bénazéraf, and Elisa Martí

17. Multiscale in vivo Imaging of Collective Cell Migration in Drosophila Embryos

Part V: Ex Vivo/InVitro Methods to Study EMT/MEt

18. Methods to Generate Tube Micropatterns for Epithelial Morphogenetic Analyses and Tissue Engineering

19. CAFs and Cancer Cells Co-Migration in 3D Spheroid Invasion Assay

Sefora Conti, Takuya Kato, Danielle Park, Erik Sahai, Xavier Trepat, Anna Labernadie

Epithelial-mesenchymal Transition

Nadège Gouignard, Christian Rouvière, and Eric Theveneau

21. Xenopus Deep Cell Aggregates: a 3D Tissue Model for Mesenchymal-To-Epithelial Transition

Hye Young Kimand Lance A. Davidson

22. Molecular Tension Microscopy of E-cadherin during Epithelial-Mesenchymal Transition

Helena Canever, Pietro Salvatore Carollo, Romain Fleurisson, Philippe P. Girard, and Nicolas Borghi

Part VI: Methods to Study Epithelial- Mesenhymal Plasticity at the Single Cell, Subcellular, and Molecular Level

23. Methodologies for Following EMT in vivo at Single Cell Resolution

Abdull J. Massri, Geoffrey R. Schiebinger, Alejandro Berrio, Lingyu Wang, Gregory A. Wray, and  David R. McClay

24. Isolation and Identification of EMT Subtypes

Robert J. Norgard and Ben Z. Stanger

25. Studying the Metabolism of Epithelial-Mesenchymal Plasticity using the Seahorse XFe96 Extracellular Flux Analyzer

Sugandha Bhatia, Erik W Thompson, and Jennifer H Gunter

26. Inducing Sequential Cycles of Epithelial-Mesenchymal and Mesenchymal-Epithelial Transitions in Mammary Epithelial Cells

Cecile Davaine, Eva Hadadi, William Taylor, Annelise Bennaceur-Griscelli, and Hervé Acloque

27. Analysis of cellular EMT States using Molecular Biology and High Resolution FISH Labeling

Noémie Kempf, Fatima Moutahir, Isabelle Goiffon, Sylvain Cantaloube, Kerstin Bystricky, and Anne-Claire Lavigne

28. Mathematical Modeling of Plasticity and Heterogeneity in EMT

Shubham Tripathi, Jianhua Xing, Herbert Levine, and Mohit Kumar Jolly

29. Control of Cell Migration Using Optogenetics

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts