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Phalloidin Staining for F-Actin in Hepatic Stellate Cells

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Hepatic Stellate Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2669))

Abstract

During the development of liver fibrosis, hepatic stellate cells undergo a transition from a quiescent phenotype into a proliferative, fibrogenic, and contractile, α-smooth muscle actin-positive myofibroblast. These cells acquire properties that are strongly associated with the reorganization of the actin cytoskeleton. Actin possesses a unique ability to polymerize into filamentous actin (F-actin) form its monomeric globular state (G-actin). F-actin can form robust actin bundles and cytoskeletal networks by interacting with a number of actin-binding proteins that provide important mechanical and structural support for a multitude of cellular processes including intracellular transport, cell motility, polarity, cell shape, gene regulation, and signal transduction. Therefore, stains with actin-specific antibodies and phalloidin conjugates for actin staining are widely used to visualize actin structures in myofibroblasts. Here we present an optimized protocol for F-actin staining for hepatic stellate cells using a fluorescent phalloidin.

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Acknowledgments

Work from Dr. Weiskirchen’s laboratory is supported by grants from the German Research Foundation (WE2554/13-1, WE2554/15-1, WE2554/17-1) and a grant from the Interdisciplinary Centre for Clinical Research within the faculty of Medicine at the RWTH Aachen University (grant PTD 1-5). The authors are grateful to Scott L. Friedman, Marcus Rojkind, and Patrick Sauvant for providing the established rat hepatic stellate cell lines HSC-T6, CFSC-2G, and PAV-1. We also thank Thomas Bauder for providing an image of the green death cap Amanita phalloides.

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Authors and Affiliations

  1. Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry (IFMPEGKC), RWTH University Hospital Aachen, Aachen, Germany

    Sarah K. Schröder, Carmen G. Tag & Sabine Weiskirchen

  2. Institut für Molekulare Pathobiochemie, Experimentelle Gentherapie und Klinische Chemie (IFMPEGKC), Universitätsklinikum Aachen AöR, Aachen, Germany

    Ralf Weiskirchen

Authors
  1. Sarah K. Schröder
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  2. Carmen G. Tag
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  3. Sabine Weiskirchen
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  4. Ralf Weiskirchen
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Corresponding authors

Correspondence to Sarah K. Schröder or Ralf Weiskirchen .

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Editors and Affiliations

  1. Institut für Molekulare Pathobiochemie, Experimentelle Gentherapie und Klinische Chemie (IFMPEGKC), Universitätsklinikum Aachen AöR, Aachen, Germany

    Ralf Weiskirchen

  2. Division of Liver Diseases, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Scott L. Friedman

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Schröder, S.K., Tag, C.G., Weiskirchen, S., Weiskirchen, R. (2023). Phalloidin Staining for F-Actin in Hepatic Stellate Cells. In: Weiskirchen, R., Friedman, S.L. (eds) Hepatic Stellate Cells. Methods in Molecular Biology, vol 2669. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3207-9_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3207-9_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3206-2

  • Online ISBN: 978-1-0716-3207-9

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