Evaluation of Nrf2 with Exposure to Nanoparticles

Methods Mol Biol. 2019:1894:229-246. doi: 10.1007/978-1-4939-8916-4_13.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / instrumentation
  • Blotting, Western / methods
  • Cells, Cultured
  • Cycloheximide / pharmacology
  • DNA Footprinting / instrumentation
  • DNA Footprinting / methods
  • Electrophoretic Mobility Shift Assay / instrumentation
  • Electrophoretic Mobility Shift Assay / methods
  • Hydroquinones / pharmacology
  • Microscopy, Confocal
  • NF-E2-Related Factor 2 / analysis*
  • NF-E2-Related Factor 2 / antagonists & inhibitors
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism
  • Nanoparticles / toxicity*
  • Oxidation-Reduction / drug effects
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology
  • RNA, Messenger / isolation & purification
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction / instrumentation
  • Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

  • Hydroquinones
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, rat
  • RNA, Messenger
  • Cycloheximide
  • 2-tert-butylhydroquinone