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Detection of SUN1 Splicing Variants at the mRNA and Protein Levels in Cancer

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The LINC Complex

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1840))

Abstract

The linker of nucleoskeleton and cytoskeleton (LINC) complex, containing the proteins SUN and nesprin, is the fundamental structural unit of the nuclear envelope. The neoplastic-based regulation of the LINC complex in cancer tissues has become increasingly recognized in recent years, including the altered expression, somatic mutation, and methylation of genes. However, precisely how mutations and deregulated expression of the LINC complex contribute to the pathogenic mechanisms of tumorigenesis remain to be elucidated, mainly because of several technical difficulties. First, both the SUN and SYNE (encoding nesprin) genes give rise to a vast number of splicing variants. Second, immunoprecipitation experiments of endogenous SUN and nesprin proteins are difficult owing to the lack of suitable reagents as well as the limited solubility of these proteins in mild extraction conditions. Here, we describe three protocols to investigate these aspects: (1) immunohistochemistry to determine the expression levels and localization of the LINC complex in cancer tissue, (2) detection of SUN1 splicing variants at the mRNA level, and (3) detection of SUN1 splicing variants and binding partners at the protein level.

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Acknowledgment

We thank Ms Yu Nishioka and Ms Junko Imada for technical assistance. This work is supported by the Education and Research Grant Program of Ehime Prefectural University of Health Science to MH.

Author information

Author notes

  1. Nariaki Matsuura

    Present address: Osaka International Cancer Institute, Osaka, Japan

  2. Miki Hieda

    Present address: Graduate School of Health Sciences, Ehime Prefectural University of Health Sciences, Iyo, Ehime, Japan

Authors and Affiliations

  1. Graduate School of Medicine and Health Science, Osaka University, Osaka, Japan

    Ayaka Matsumoto, Nariaki Matsuura & Miki Hieda

Authors
  1. Ayaka Matsumoto
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  2. Nariaki Matsuura
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  3. Miki Hieda
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Editor information

Editors and Affiliations

  1. Department of Pathology and Cell Biology, Columbia University, New York, New York, USA

    Gregg G. Gundersen

  2. Department of Medicine and Department of Pathology and Cell Biology, Columbia University, New York, New York, USA

    Howard J. Worman

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Matsumoto, A., Matsuura, N., Hieda, M. (2018). Detection of SUN1 Splicing Variants at the mRNA and Protein Levels in Cancer. In: Gundersen, G., Worman, H. (eds) The LINC Complex. Methods in Molecular Biology, vol 1840. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8691-0_21

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  • DOI: https://doi.org/10.1007/978-1-4939-8691-0_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8690-3

  • Online ISBN: 978-1-4939-8691-0

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