Abstract
Transcription factor Nrf2, nuclear factor (erythroid-derived 2)-like 2, is considered a master regulator of redox homeostasis and plays a central role in antioxidant and anti-inflammatory defence. It has been largely reported that oxidative stress is implicated in nanoparticle-induced toxicity with the involvement of Nrf2. Several basic methods for Nrf2 evaluation with exposure to nanoparticles are described in this chapter including real-time reverse transcription-polymerase chain reaction (RT-PCR), western blotting, immunofluorescence staining, electrophoretic mobility shift assay, DNase I footprinting, dimethylsulfate footprinting, protein pulse-chase analysis, and tert-butylhydroquinone treatment.
Keywords:
Antioxidant response element (ARE); EMSA; Footprinting; Nanoparticles; Nrf2; Pulse chase analysis; t-BHQ.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Blotting, Western / instrumentation
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Blotting, Western / methods
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Cells, Cultured
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Cycloheximide / pharmacology
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DNA Footprinting / instrumentation
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DNA Footprinting / methods
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Electrophoretic Mobility Shift Assay / instrumentation
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Electrophoretic Mobility Shift Assay / methods
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Hydroquinones / pharmacology
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Microscopy, Confocal
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NF-E2-Related Factor 2 / analysis*
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NF-E2-Related Factor 2 / antagonists & inhibitors
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NF-E2-Related Factor 2 / genetics
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NF-E2-Related Factor 2 / metabolism
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Nanoparticles / toxicity*
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Oxidation-Reduction / drug effects
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Oxidative Stress / drug effects
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Oxidative Stress / physiology
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RNA, Messenger / isolation & purification
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Rats
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Reverse Transcriptase Polymerase Chain Reaction / instrumentation
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Reverse Transcriptase Polymerase Chain Reaction / methods
Substances
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Hydroquinones
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NF-E2-Related Factor 2
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Nfe2l2 protein, rat
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RNA, Messenger
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Cycloheximide
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2-tert-butylhydroquinone