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Molecular Toxicology Protocols pp 531–555Cite as

Methods for Establishing and Using a Stable Cell Line Expressing Both Gaussia Luciferase and Firefly Luciferase to Screen for Endoplasmic Reticulum Stress

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2102))

Abstract

Endoplasmic reticulum (ER) stress is one of the major mechanisms underlying the etiology of multiple diseases and drug-induced toxicity. Gaussia luciferase (Gluc) is a naturally secreted protein that has been used as a reporter for the secretory pathway of ER to enable efficient and real-time monitoring of the ER function. The Gluc assay has been widely used and optimized in various labs. In this chapter, we provide an example of the application of the Gluc assay by establishing a stable cell line expressing both Gluc and firefly luciferase (Fluc) to study ER stress in liver cells. We describe the detailed procedures used in our laboratory for Gluc- and Fluc-containing lentivirus production and titration, for establishing a HepG2-based stable cell line through lentivirus transduction and the validation process. In addition, we provide an example of using the established stable cell line to investigate ER stress.

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Acknowledgment

Use of trade names is for informational purposes only and in no way implies endorsement by the U.S. Food and Drug Administration (FDA). The views presented in this paper do not necessarily reflect those of the U.S. FDA.

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Authors and Affiliations

  1. Division of Biochemical Toxicology, National Center for Toxicological Research (NCTR)/U.S. FDA, Jefferson, AR, USA

    Zhen Ren, Si Chen & Lei Guo

Authors
  1. Zhen Ren
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  2. Si Chen
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  3. Lei Guo
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Corresponding author

Correspondence to Lei Guo .

Editor information

Editors and Affiliations

  1. Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA, USA

    Phouthone Keohavong

  2. Department of Environmental Toxicology, The Institute of Environmental and Human Health (TIEHH), Texas Tech University, Lubbock, TX, USA

    Kamaleshwar P. Singh

  3. Department of Occupational and Environmental Health Sciences, School of Public Health West Virginia University, Morgantown, WV, USA

    Weimin Gao

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Ren, Z., Chen, S., Guo, L. (2020). Methods for Establishing and Using a Stable Cell Line Expressing Both Gaussia Luciferase and Firefly Luciferase to Screen for Endoplasmic Reticulum Stress. In: Keohavong, P., Singh, K., Gao, W. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 2102. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0223-2_29

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  • DOI: https://doi.org/10.1007/978-1-0716-0223-2_29

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0222-5

  • Online ISBN: 978-1-0716-0223-2

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