Epidermal Cells

Volume 289 of the series Methods in Molecular Biology™ pp 147-156

Analysis of E2F Factors During Epidermal Differentiation

  • Wing Y. ChangAffiliated withDepartment of Physiology and Pharmacology, University of Western Ontario
  • , Lina DagninoAffiliated withDepartments of Physiology and Pharmacology and of Pediatrics, University of Western Ontario

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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.

Key Words

E2F transcription factors epidermis keratinocytes proliferation differentiation