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Evaluation of Nrf2 with Exposure to Nanoparticles
Fuli Zheng1,2, Huangyuan Li3,4
1Fujian Provincial Key Laboratory of Environmental Factors and Cancer, School of Public Health, Fujian Medical University, Fuzhou, P. R. China.
Methods in Molecular Biology (Clifton, N.J.)
|December 15, 2018
Summary
This chapter details methods for evaluating the role of transcription factor Nrf2 in nanoparticle toxicity. It covers techniques like RT-PCR and western blotting to assess Nrf2
Area of Science:
- Biochemistry
- Toxicology
- Molecular Biology
Background:
- Transcription factor Nrf2 (nuclear factor erythroid-derived 2-like 2) is a key regulator of cellular redox homeostasis.
- Oxidative stress is a known factor in nanoparticle-induced toxicity, often involving Nrf2.
- Understanding Nrf2's role is crucial for assessing nanoparticle safety.
Purpose of the Study:
- To outline methods for evaluating Nrf2 activity following nanoparticle exposure.
- To provide a resource for researchers studying nanoparticle toxicology and Nrf2 pathways.
Main Methods:
- Real-time reverse transcription-polymerase chain reaction (RT-PCR) for Nrf2 gene expression.
- Western blotting to assess Nrf2 protein levels.
- Immunofluorescence staining for Nrf2 localization.
- Electrophoretic mobility shift assay (EMSA) and footprinting techniques for DNA binding.
- Protein pulse-chase analysis and tert-butylhydroquinone treatment for functional assessment.
Main Results:
- The chapter describes multiple techniques to measure Nrf2's response to nanoparticles.
- These methods allow for comprehensive evaluation of Nrf2's involvement in nanoparticle toxicity.
- Specific protocols for gene expression, protein levels, DNA binding, and functional assays are presented.
Conclusions:
- Accurate evaluation of Nrf2 is essential for understanding nanoparticle toxicity.
- A range of molecular and biochemical methods are available for Nrf2 assessment.
- This chapter serves as a guide to these critical evaluation techniques.
Keywords:
Antioxidant response element (ARE)EMSAFootprintingNanoparticlesNrf2Pulse chase analysist-BHQMore Related Videos
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