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High-Resolution Imaging of STIM/Orai Subcellular Localization Using Array Confocal Laser Scanning Microscopy

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The CRAC Channel

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1843))

Abstract

The expression of chimeras that consist of a fluorescent protein (FP) conjugated with a protein of interest provides the ability to visualize, track, and quantify the subcellular localization and dynamics of specific proteins in biological samples. Array confocal laser scanning microscopy is an eminently suitable technique for live-cell imaging of FP-tagged fusion proteins. Here, we describe real-time monitoring of the subcellular dynamics of the stromal-interacting molecule 1 (STIM1) and Orai1, the key protagonists of store-operated Ca2+ entry (SOCE) under resting conditions, and upon Ca2+ mobilization from the endoplasmic reticulum (ER).

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Acknowledgments

We thank Sandra Blass and Anna Schreilechner for their excellent technical assistance and Dr. C.J.S. Edgell (University of North Carolina, Chapel Hill, NC, USA) for the EA.hy926 cells. This work was supported by the Austrian Science Funds (FWF) with the project number P 28529-B27 and also sponsoring the DKplus Metabolic and Cardiovascular Disease (W1226-B18) of the Medical University of Graz. A.T.D., B.G., and E.E. was/are a fellow/fellows of the Doctoral College “Metabolic and Cardiovascular Disease” at the Medical University of Graz and was/are funded by the FWF (A.T.D.), Nikon Austria (B.G.), or BioTechMed (E.E.) (both within the Nikon-Center of Excellence, Graz) within the Doctoral College “Metabolic and Cardiovascular Disease” (FWF W 1226-B18, DKplus Metabolic and Cardiovascular Disease).

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Authors and Affiliations

  1. Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria

    Andras T. Deak, Benjamin Gottschalk, Emrah Eroglu, Rene Rost, Markus Waldeck-Weiermair, Wolfgang F. Graier & Roland Malli

  2. Clinical Division of Nephrology, Department of Internal Medicine, Medical University of Graz, Graz, Austria

    Andras T. Deak

Authors
  1. Andras T. Deak
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  2. Benjamin Gottschalk
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  3. Emrah Eroglu
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  4. Rene Rost
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  5. Markus Waldeck-Weiermair
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  6. Wolfgang F. Graier
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  7. Roland Malli
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Corresponding author

Correspondence to Roland Malli .

Editor information

Editors and Affiliations

  1. IRSET Inserm UMR 1085/ UR1, Rennes, France

    Aubin Penna

  2. CNRS ERL 7003, Laboratoire STIM, Poitiers, France

    Bruno Constantin

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Deak, A.T. et al. (2018). High-Resolution Imaging of STIM/Orai Subcellular Localization Using Array Confocal Laser Scanning Microscopy. In: Penna, A., Constantin, B. (eds) The CRAC Channel. Methods in Molecular Biology, vol 1843. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8704-7_15

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  • DOI: https://doi.org/10.1007/978-1-4939-8704-7_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8702-3

  • Online ISBN: 978-1-4939-8704-7

  • eBook Packages: Springer Protocols

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