17. Application of Electrophoretic Mobility Shift Assay and Chromatin Immunoprecipitation in the Study of Transcription in Adipose
Cells
| Abstract |
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Chromatin, long thought to be no more than a scaffold supporting DNA compaction inside the cell nucleus, has emerged in the
last few years as a major regulatory element involved in the control of gene expression both acutely during interphase and
programmatically throughout complex processes of development and differentiation. Adipogenesis is the result of an intertwined
network of transcription factors and coregulators with chromatin-modifying activities and offers an excellent model for the
study of transcriptional regulation. In this regard, electrophoretic mobility shift assay and immunoprecipitation of chromatin
are complementary methods that can be used to study the binding of nuclear proteins to DNA and to characterize how these proteins
interact with and modify chromatin to regulate gene expression and, more globally, cell differentiation. This chapter provides
some strategies to perform these two assays using 3T3-L1 cells and rodent primary preadipocytes and adipocytes.
Affiliation(s): (3) Endocrinology and Nutrition Unit, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) Hospital Clínic, Universitat de Barcelona, Barcelona, Spain
Book Title: Adipose Tissue Protocols
Series: Methods in Molecular Biology | Volume: 456 | Pub. Date: Jun-01-2008 | Page Range: 231-247 | DOI: 10.1007/978-1-59745-245-8_17
Subject: Molecular Medicine
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