Skip to main content
SpringerLink
Log in

Analysis of E2F Factors During Epidermal Differentiation

  • Protocol
Epidermal Cells

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 289))

Abstract

The multigene E2F family of transcription factors is central in the control of cell cycle progression. The expression and activity of E2F proteins is tightly regulated transcriptionally and posttranslationally as a function of the proliferation and differentiation status of the cell. In this chapter, we review protocols designed to determine E2F mRNA abundance in tissues by in situ hybridization techniques. The ability to culture primary epidermal keratinocytes and maintain them as either undifferentiated or terminally differentiated cells allows the biochemical and molecular characterization of changes in E2F expression and activity. Thus, we also discuss in detail methods to analyze E2F protein abundance by immunoblot and their ability to bind DNA in cultured cells using electrophoretic mobility shift assays.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Dotto, G. P. (1999) Signal transduction pathways controlling the switch between keratinocyte growth and differentiation. Crit. Rev. Oral Biol. Med. 10, 442–457.

    Article  PubMed  CAS  Google Scholar 

  2. D’souza, S. J. A., Pajak, A., Balazsi, K., and Dagnino, L. (2001) Ca2+ and BMP-6 signalling regulate E2F during epidermal keratinocyte differentiation. J. Biol. Chem. 276, 23,531–23,538.

    Article  CAS  Google Scholar 

  3. Dagnino, L., Fry, C. J., Bartley, S. M., Farnham, P., Gallie, B. L., and Phillips, R. A. (1997) Expression patterns of the E2F family of transcription factors during mouse nervous system development. Mech. Dev. 66, 13–25.

    Article  PubMed  CAS  Google Scholar 

  4. Dagnino, L., Fry, C. J., Bartley, S. M., Farnham, P., Gallie, B. L., and Phillips, R. A. (1997) Expression patterns of the E2F family of transcription factors during murine epithelial development. Cell Growth Differ. 8, 553–563.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada

    Wing Y. Chang

  2. Departments of Physiology and Pharmacology and of Pediatrics, University of Western Ontario, London, Ontario, Canada

    Lina Dagnino

Authors
  1. Wing Y. Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lina Dagnino
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Ottawa Health Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Humana Press Inc.

About this protocol

Cite this protocol

Chang, W.Y., Dagnino, L. (2005). Analysis of E2F Factors During Epidermal Differentiation. In: Turksen, K. (eds) Epidermal Cells. Methods in Molecular Biology™, vol 289. Humana Press. https://doi.org/10.1385/1-59259-830-7:147

Download citation

  • DOI: https://doi.org/10.1385/1-59259-830-7:147

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-267-4

  • Online ISBN: 978-1-59259-830-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation