Analysis of focal adhesions and cytoskeleton by custom microarray

Matthew J. Dalby, S. J. Yarwood

    Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

    9 Citations (Scopus)

    Abstract

    Focal adhesions and the cell cytoskeleton (intermediate filaments, microfilaments, microtubules) are involved in mechanotransduction-both direct (transduction of mechanical forces to the nucleus) and indirect (transduction of chemical signaling cascades to the nucleus). Thus, observation of changes in focal adhesion and cytoskeletal organization can be invaluable in research such as drug treatments and medical material testing in vitro. Here we describe how to stain human fibroblasts for vinculin (located to focal adhesions), actin (microfilaments), tubulin (microtubules), and vimentin (intermediate filaments) and how to perform custom microarray experiments. Comparative analysis of the immunofluorescence and array data should allow the researcher to build up a global picture of changes to both direct and indirect mechanotransduction through the cytoskeleton from focal adhesions.

    Original languageEnglish
    Title of host publicationAdhesion Protein Protocols
    EditorsAmanda S. Coutts
    PublisherHumana Press
    Pages121-134
    Number of pages14
    ISBN (Electronic)978-1-59745-353-0
    ISBN (Print)978-1-58829-533-0
    DOIs
    Publication statusPublished - 2007

    Publication series

    NameMethods in Molecular Biology
    PublisherHumana Press
    Volume370
    ISSN (Print)1064-3745

    Keywords

    • Actin Cytoskeleton
    • Actins
    • Cell Line
    • Cells, Cultured
    • Cluster Analysis
    • Cytoskeleton
    • Fibroblasts
    • Fluorescent Antibody Technique
    • Focal Adhesions
    • Gene Expression Profiling
    • Humans
    • Immunohistochemistry
    • Intermediate Filaments
    • Microtubules
    • Oligonucleotide Array Sequence Analysis
    • Tubulin
    • Vimentin
    • Vinculin

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