qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathway: Summary
Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration. The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of oxidative stress. The CellSensor ARE-bla HepG2 cell line (Invitrogen) can be used for analyzing the Nrf2/antioxidant response signaling pathway. Nrf2 (NF-E2-related factor more ..
BioActive Compounds: 995
Depositor Specified Assays
U.S. Tox21 Program
National Center for Advancing Translational Sciences [NCATS]
NIH Chemical Genomics Center [NCGC]
U.S. Environmental Protection Agency [EPA]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]
U.S. Food and Drug Administration [FDA]
Tox21 Assay Overview:
Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration. The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of oxidative stress. The CellSensor ARE-bla HepG2 cell line (Invitrogen) can be used for analyzing the Nrf2/antioxidant response signaling pathway. Nrf2 (NF-E2-related factor 2) and Nrf1 are transcription factors that bind to AREs and activate these genes. The CellSensor ARE-bla Hep G2 cell line contains a beta-lactamase reporter gene under control of the Antioxidant Response Element (ARE) stably integrated into HepG2 cells. The activation of the report gene under culture conditions can be detected by fluorescence intensity. This cell line has been used to screen the Tox21 10K compound library to identify agonists that induce oxidative stress. The cytotoxicity of the Tox21 compound library against the ARE-bla cell line was tested in parallel by measuring the cell viability using CellTiter-Glo assay (Promega, Madison, WI) in the same wells. The compounds were also tested for auto fluorescence that may interfere with the biological target readout resulting in potential false positives and/or negatives.
Please refer to other AIDs 743202, 743203, 720687, 720685, 720678 and 720681, for detailed assay protocols.
This summary is written for the purposes of summarizing the compound activities from the project combining the results from both the ARE agonist mode assay (AID 743202), cell viability counter screen ( 743203), and auto fluorescence counter screens (AIDs 720687, 720685, 720678 and 720681). For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Potency and efficacy were used for determining relative score. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 5 and 30 determined by phenotype.
Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods. Alternative analysis methods and interpretations of the data are available at EPA (http://actor.epa.gov) and NTP (http://tools.niehs.nih.gov/cebs3/ui/).
* Activity Concentration.
Data Table (Concise)