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Lifeact: a versatile marker to visualize F-actin

Nature Methods volume 5, pages 605–607 (2008)Cite this article

Abstract

Live imaging of the actin cytoskeleton is crucial for the study of many fundamental biological processes, but current approaches to visualize actin have several limitations. Here we describe Lifeact, a 17-amino-acid peptide, which stained filamentous actin (F-actin) structures in eukaryotic cells and tissues. Lifeact did not interfere with actin dynamics in vitro and in vivo and in its chemically modified peptide form allowed visualization of actin dynamics in nontransfectable cells.

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Figure 1: Identification and biochemical characterization of Lifeact.
Figure 2: Characterization of Lifeact-GFP in vivo.
Figure 3: F-Lifeact staining in fixed and living samples.

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Acknowledgements

We thank S. Uebel and the core facility of the Max Planck Institute of Biochemistry for peptide synthesis and labeling, and S. Cremer and S. Wedlich for helpful comments on the manuscript. We obtained latrunculin A from P. Crews (University of California Santa Cruz). This study was supported by funds from the Max Planck Society (R.W.-S.), German Research Foundation SI1323/1-1 (M.S.) and the Alexander von Humboldt Stiftung (Z.W.).

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Author notes

  1. Julia Riedl and Alvaro H Crevenna: These authors contributed equally to this work.

Authors and Affiliations

  1. Max Planck Institute of Biochemistry, Independent Junior Research Group Cellular Dynamics and Cell Patterning, Am Klopferspitz 18, Martinsried, 82152, Germany

    Julia Riedl, Alvaro H Crevenna, Jerry Haochen Yu & Roland Wedlich-Soldner

  2. Department of Neuroimmunology, Max Planck Institute of Neurobiology, Am Klopferspitz 18, Martinsried, 82152, Germany

    Kai Kessenbrock & Dieter Jenne

  3. Max Planck Institute of Neurobiology, Independent Junior Research Group Axonal Growth and Regeneration, Am Klopferspitz 18, Martinsried, 82152, Germany

    Dorothee Neukirchen & Frank Bradke

  4. Department of Structural Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, Martinsried, 82152, Germany

    Michal Bista & Tad A Holak

  5. Department of Anatomy and The Biomedical Sciences Program, University of California, San Francisco, San Francisco, 513 Parnassus Avenue, 94143, California, USA

    Zena Werb

  6. Department of Molecular Medicine, Max Planck Institute of Biochemistry, Am Klopferspitz 18, Martinsried, 82152, Germany

    Michael Sixt

Authors
  1. Julia Riedl
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Corresponding authors

Correspondence to Michael Sixt or Roland Wedlich-Soldner.

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Competing interests

J.R., A.H.C., M.S. and R.W.-S. have submitted a patent application (07 023 084.2; Peptide for determining actin structures in living cells) to the EU patent office.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1, Supplementary Methods (PDF 193 kb)

Supplementary Movie 1

Actin dynamics in a mouse embryonic fibroblast. Images were acquired at 9 s/frame and are played back at 10 frames/s. Time stamp in minutes and seconds. (MOV 1655 kb)

Supplementary Movie 2

Actin dynamics in a chemotactic mouse dendritic cell. Images were acquired at 3 s/frame and are played back at 10 frames/s. Time stamp in minutes and seconds. (MOV 3004 kb)

Supplementary Movie 3

Dynamics of filopodia in a hippocampal neuron. Images were acquired at 2 s/frame and are played back at 10 frames/s. Time stamp in minutes and seconds. (MOV 820 kb)

Supplementary Movie 4

Cortical actin dynamics in the cell body of a hippocampal neuron. Images were acquired at 2 s/frame and are played back at 10 frames/s. Time stamp in minutes and seconds. (MOV 3804 kb)

Supplementary Movie 5

Cytokinetic ring formation during mitosis of MDCK cells. Images were acquired at 2 min/frame and are played back with 10 frames/s. Time stamp in hours and minutes. (MOV 238 kb)

Supplementary Movie 6

Spreading of a primary human neutrophil on a cover petri dish coated with immune complexes. Images were acquired at 1 s/frame and are played back at 10 frames/s. Time stamp in minutes and seconds. (MOV 1832 kb)

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Riedl, J., Crevenna, A., Kessenbrock, K. et al. Lifeact: a versatile marker to visualize F-actin. Nat Methods 5, 605–607 (2008). https://doi.org/10.1038/nmeth.1220

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