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BioAssay: AID 720653

qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathway measured by Nrf2-dependant transcriptional activity - cell viability counter screen

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 ..
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 Tested Compounds
 Tested Compounds
All(2329)
 
 
Active(56)
 
 
Inactive(2200)
 
 
Inconclusive(108)
 
 
 Tested Substances
 Tested Substances
All(2864)
 
 
Active(62)
 
 
Inactive(2686)
 
 
Inconclusive(116)
 
 
 Related BioAssays
 Related BioAssays
AID: 720653
Data Source: NCGC (ARE853)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2013-10-17

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 56
Related Experiments
AIDNameTypeComment
651741qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathwayConfirmatorydepositor-specified cross reference
Description:
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]

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 HepG2 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. The cytotoxicity of the compounds against the ARE-bla HepG2 cell line was quantified by measuring the intracellular ATP content.
Protocol
Assay Protocol Summary:
ARE-bla HepG2 cells were dispensed at 2,000 cells/5 ul/well in 1536-well black wall/clear bottom plates using a Multidrop Combi (ThermoFisher Scientific, Waltham, MA) dispenser. After the assay plates were incubated at 37C/5% CO2 for 5 hrs, 23 nL of compounds dissolved in DMSO, positive controls or DMSO only was transferred to the assay plate by a pintool work station (Kalypsys, San Diego, CA). The plates were incubated at 37C/5% CO2 for 16 hrs. After 1 uL of LiveBLAzer B/G FRET substrate was added using a Flying Reagent Dispenser (FRD) (Aurora Discovery, San Diego, CA), the assay plates were incubated at room temperature for 2 hrs. The fluorescence intensity was measured by an Envision (PerkinElmer, Shelton, CT) plate reader. Cell viability using CellTiter Glo viability assay (Promega, Madison, WI) was also measured by adding 4 ul/well of CellTiter-Glo reagent into the same assay plate using a FRD. After 30 min incubation at room temperature, the luminescence intensity in the plates was measured using a ViewLux (PerkinElmer) plate reader.
Comment
Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: HEPG2
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Phenotype-Replicate_1Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency-Replicate_1*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3Efficacy-Replicate_1Maximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis Comment-Replicate_1Annotation/notes on a particular compound's data or its analysis.String
5Activity_Score-Replicate_1Activity score.Integer
6Curve_Description-Replicate_1A description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50-Replicate_1The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlope-Replicate_1The Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2-Replicate_1R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivity-Replicate_1The asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivity-Replicate_1Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClass-Replicate_1Numerical encoding of curve description for the fitted Hill equation.Float
13Excluded_Points-Replicate_1Which dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_Response-Replicate_1Maximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0005213331 uM-Replicate_1 (0.000521333μM**)% Activity at given concentration.Float%
16Activity at 0.00156 uM-Replicate_1 (0.0015582μM**)% Activity at given concentration.Float%
17Activity at 0.00466 uM-Replicate_1 (0.00465609μM**)% Activity at given concentration.Float%
18Activity at 0.00590 uM-Replicate_1 (0.00589862μM**)% Activity at given concentration.Float%
19Activity at 0.014 uM-Replicate_1 (0.0140533μM**)% Activity at given concentration.Float%
20Activity at 0.042 uM-Replicate_1 (0.0417583μM**)% Activity at given concentration.Float%
21Activity at 0.068 uM-Replicate_1 (0.0684607μM**)% Activity at given concentration.Float%
22Activity at 0.127 uM-Replicate_1 (0.126926μM**)% Activity at given concentration.Float%
23Activity at 0.378 uM-Replicate_1 (0.378213μM**)% Activity at given concentration.Float%
24Activity at 0.768 uM-Replicate_1 (0.767702μM**)% Activity at given concentration.Float%
25Activity at 1.156 uM-Replicate_1 (1.15581μM**)% Activity at given concentration.Float%
26Activity at 3.420 uM-Replicate_1 (3.41952μM**)% Activity at given concentration.Float%
27Activity at 5.843 uM-Replicate_1 (5.84271μM**)% Activity at given concentration.Float%
28Activity at 10.21 uM-Replicate_1 (10.2119μM**)% Activity at given concentration.Float%
29Activity at 30.24 uM-Replicate_1 (30.2438μM**)% Activity at given concentration.Float%
30Activity at 43.68 uM-Replicate_1 (43.6833μM**)% Activity at given concentration.Float%
31Activity at 92.27 uM-Replicate_1 (92.2654μM**)% Activity at given concentration.Float%
32Activity at 218.9 uM-Replicate_1 (218.934μM**)% Activity at given concentration.Float%
33Activity at 489.4 uM-Replicate_1 (489.435μM**)% Activity at given concentration.Float%
34Activity at 1094.5 uM-Replicate_1 (1094.55μM**)% Activity at given concentration.Float%
35Activity at 2447.4 uM-Replicate_1 (2447.39μM**)% Activity at given concentration.Float%
36Activity at 5811.1 uM-Replicate_1 (5811.06μM**)% Activity at given concentration.Float%
37Activity at 12995.4 uM-Replicate_1 (12995.4μM**)% Activity at given concentration.Float%
38Activity at 29059.9 uM-Replicate_1 (29059.9μM**)% Activity at given concentration.Float%
39Activity at 64977.0 uM-Replicate_1 (64977μM**)% Activity at given concentration.Float%
40Compound QC-Replicate_1NCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String
41Phenotype-Replicate_2Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
42Potency-Replicate_2Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
43Efficacy-Replicate_2Maximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
44Analysis Comment-Replicate_2Annotation/notes on a particular compound's data or its analysis.String
45Activity_Score-Replicate_2Activity score.Integer
46Curve_Description-Replicate_2A description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
47Fit_LogAC50-Replicate_2The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
48Fit_HillSlope-Replicate_2The Hill slope from a fit of the data to the Hill equation.Float
49Fit_R2-Replicate_2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
50Fit_InfiniteActivity-Replicate_2The asymptotic efficacy from a fit of the data to the Hill equation.Float%
51Fit_ZeroActivity-Replicate_2Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
52Fit_CurveClass-Replicate_2Numerical encoding of curve description for the fitted Hill equation.Float
53Excluded_Points-Replicate_2Which dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
54Max_Response-Replicate_2Maximum activity observed for compound (usually at highest concentration tested).Float%
55Activity at 0.0005213331 uM-Replicate_2 (0.000521333μM**)% Activity at given concentration.Float%
56Activity at 0.00156 uM-Replicate_2 (0.0015582μM**)% Activity at given concentration.Float%
57Activity at 0.00466 uM-Replicate_2 (0.00465609μM**)% Activity at given concentration.Float%
58Activity at 0.00590 uM-Replicate_2 (0.00589862μM**)% Activity at given concentration.Float%
59Activity at 0.014 uM-Replicate_2 (0.0140533μM**)% Activity at given concentration.Float%
60Activity at 0.042 uM-Replicate_2 (0.0417583μM**)% Activity at given concentration.Float%
61Activity at 0.068 uM-Replicate_2 (0.0684607μM**)% Activity at given concentration.Float%
62Activity at 0.127 uM-Replicate_2 (0.126926μM**)% Activity at given concentration.Float%
63Activity at 0.378 uM-Replicate_2 (0.378213μM**)% Activity at given concentration.Float%
64Activity at 0.768 uM-Replicate_2 (0.767702μM**)% Activity at given concentration.Float%
65Activity at 1.156 uM-Replicate_2 (1.15581μM**)% Activity at given concentration.Float%
66Activity at 3.420 uM-Replicate_2 (3.41952μM**)% Activity at given concentration.Float%
67Activity at 5.843 uM-Replicate_2 (5.84271μM**)% Activity at given concentration.Float%
68Activity at 10.21 uM-Replicate_2 (10.2119μM**)% Activity at given concentration.Float%
69Activity at 30.24 uM-Replicate_2 (30.2438μM**)% Activity at given concentration.Float%
70Activity at 43.68 uM-Replicate_2 (43.6833μM**)% Activity at given concentration.Float%
71Activity at 92.27 uM-Replicate_2 (92.2654μM**)% Activity at given concentration.Float%
72Activity at 218.9 uM-Replicate_2 (218.934μM**)% Activity at given concentration.Float%
73Activity at 489.4 uM-Replicate_2 (489.435μM**)% Activity at given concentration.Float%
74Activity at 1094.5 uM-Replicate_2 (1094.55μM**)% Activity at given concentration.Float%
75Activity at 2447.4 uM-Replicate_2 (2447.39μM**)% Activity at given concentration.Float%
76Activity at 5811.1 uM-Replicate_2 (5811.06μM**)% Activity at given concentration.Float%
77Activity at 12995.4 uM-Replicate_2 (12995.4μM**)% Activity at given concentration.Float%
78Activity at 29059.9 uM-Replicate_2 (29059.9μM**)% Activity at given concentration.Float%
79Activity at 64977.0 uM-Replicate_2 (64977μM**)% Activity at given concentration.Float%
80Phenotype-Replicate_3Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
81Potency-Replicate_3Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
82Efficacy-Replicate_3Maximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
83Analysis Comment-Replicate_3Annotation/notes on a particular compound's data or its analysis.String
84Activity_Score-Replicate_3Activity score.Integer
85Curve_Description-Replicate_3A description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
86Fit_LogAC50-Replicate_3The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
87Fit_HillSlope-Replicate_3The Hill slope from a fit of the data to the Hill equation.Float
88Fit_R2-Replicate_3R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
89Fit_InfiniteActivity-Replicate_3The asymptotic efficacy from a fit of the data to the Hill equation.Float%
90Fit_ZeroActivity-Replicate_3Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
91Fit_CurveClass-Replicate_3Numerical encoding of curve description for the fitted Hill equation.Float
92Excluded_Points-Replicate_3Which dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
93Max_Response-Replicate_3Maximum activity observed for compound (usually at highest concentration tested).Float%
94Activity at 0.0005213331 uM-Replicate_3 (0.000521333μM**)% Activity at given concentration.Float%
95Activity at 0.00156 uM-Replicate_3 (0.0015582μM**)% Activity at given concentration.Float%
96Activity at 0.00466 uM-Replicate_3 (0.00465609μM**)% Activity at given concentration.Float%
97Activity at 0.00590 uM-Replicate_3 (0.00589862μM**)% Activity at given concentration.Float%
98Activity at 0.014 uM-Replicate_3 (0.0140533μM**)% Activity at given concentration.Float%
99Activity at 0.042 uM-Replicate_3 (0.0417583μM**)% Activity at given concentration.Float%
100Activity at 0.068 uM-Replicate_3 (0.0684607μM**)% Activity at given concentration.Float%
101Activity at 0.127 uM-Replicate_3 (0.126926μM**)% Activity at given concentration.Float%
102Activity at 0.378 uM-Replicate_3 (0.378213μM**)% Activity at given concentration.Float%
103Activity at 0.768 uM-Replicate_3 (0.767702μM**)% Activity at given concentration.Float%
104Activity at 1.156 uM-Replicate_3 (1.15581μM**)% Activity at given concentration.Float%
105Activity at 3.420 uM-Replicate_3 (3.41952μM**)% Activity at given concentration.Float%
106Activity at 5.843 uM-Replicate_3 (5.84271μM**)% Activity at given concentration.Float%
107Activity at 10.21 uM-Replicate_3 (10.2119μM**)% Activity at given concentration.Float%
108Activity at 30.24 uM-Replicate_3 (30.2438μM**)% Activity at given concentration.Float%
109Activity at 43.68 uM-Replicate_3 (43.6833μM**)% Activity at given concentration.Float%
110Activity at 92.27 uM-Replicate_3 (92.2654μM**)% Activity at given concentration.Float%
111Activity at 218.9 uM-Replicate_3 (218.934μM**)% Activity at given concentration.Float%
112Activity at 489.4 uM-Replicate_3 (489.435μM**)% Activity at given concentration.Float%
113Activity at 1094.5 uM-Replicate_3 (1094.55μM**)% Activity at given concentration.Float%
114Activity at 2447.4 uM-Replicate_3 (2447.39μM**)% Activity at given concentration.Float%
115Activity at 5811.1 uM-Replicate_3 (5811.06μM**)% Activity at given concentration.Float%
116Activity at 12995.4 uM-Replicate_3 (12995.4μM**)% Activity at given concentration.Float%
117Activity at 29059.9 uM-Replicate_3 (29059.9μM**)% Activity at given concentration.Float%
118Activity at 64977.0 uM-Replicate_3 (64977μM**)% Activity at given concentration.Float%
119Phenotype-Replicate_4Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
120Potency-Replicate_4Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
121Efficacy-Replicate_4Maximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
122Analysis Comment-Replicate_4Annotation/notes on a particular compound's data or its analysis.String
123Activity_Score-Replicate_4Activity score.Integer
124Curve_Description-Replicate_4A description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
125Fit_LogAC50-Replicate_4The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
126Fit_HillSlope-Replicate_4The Hill slope from a fit of the data to the Hill equation.Float
127Fit_R2-Replicate_4R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
128Fit_InfiniteActivity-Replicate_4The asymptotic efficacy from a fit of the data to the Hill equation.Float%
129Fit_ZeroActivity-Replicate_4Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
130Fit_CurveClass-Replicate_4Numerical encoding of curve description for the fitted Hill equation.Float
131Excluded_Points-Replicate_4Which dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
132Max_Response-Replicate_4Maximum activity observed for compound (usually at highest concentration tested).Float%
133Activity at 0.0005213331 uM-Replicate_4 (0.000521333μM**)% Activity at given concentration.Float%
134Activity at 0.00156 uM-Replicate_4 (0.0015582μM**)% Activity at given concentration.Float%
135Activity at 0.00466 uM-Replicate_4 (0.00465609μM**)% Activity at given concentration.Float%
136Activity at 0.00590 uM-Replicate_4 (0.00589862μM**)% Activity at given concentration.Float%
137Activity at 0.014 uM-Replicate_4 (0.0140533μM**)% Activity at given concentration.Float%
138Activity at 0.042 uM-Replicate_4 (0.0417583μM**)% Activity at given concentration.Float%
139Activity at 0.068 uM-Replicate_4 (0.0684607μM**)% Activity at given concentration.Float%
140Activity at 0.127 uM-Replicate_4 (0.126926μM**)% Activity at given concentration.Float%
141Activity at 0.378 uM-Replicate_4 (0.378213μM**)% Activity at given concentration.Float%
142Activity at 0.768 uM-Replicate_4 (0.767702μM**)% Activity at given concentration.Float%
143Activity at 1.156 uM-Replicate_4 (1.15581μM**)% Activity at given concentration.Float%
144Activity at 3.420 uM-Replicate_4 (3.41952μM**)% Activity at given concentration.Float%
145Activity at 5.843 uM-Replicate_4 (5.84271μM**)% Activity at given concentration.Float%
146Activity at 10.21 uM-Replicate_4 (10.2119μM**)% Activity at given concentration.Float%
147Activity at 30.24 uM-Replicate_4 (30.2438μM**)% Activity at given concentration.Float%
148Activity at 43.68 uM-Replicate_4 (43.6833μM**)% Activity at given concentration.Float%
149Activity at 92.27 uM-Replicate_4 (92.2654μM**)% Activity at given concentration.Float%
150Activity at 218.9 uM-Replicate_4 (218.934μM**)% Activity at given concentration.Float%
151Activity at 489.4 uM-Replicate_4 (489.435μM**)% Activity at given concentration.Float%
152Activity at 1094.5 uM-Replicate_4 (1094.55μM**)% Activity at given concentration.Float%
153Activity at 2447.4 uM-Replicate_4 (2447.39μM**)% Activity at given concentration.Float%
154Activity at 5811.1 uM-Replicate_4 (5811.06μM**)% Activity at given concentration.Float%
155Activity at 12995.4 uM-Replicate_4 (12995.4μM**)% Activity at given concentration.Float%
156Activity at 29059.9 uM-Replicate_4 (29059.9μM**)% Activity at given concentration.Float%
157Activity at 64977.0 uM-Replicate_4 (64977μM**)% Activity at given concentration.Float%

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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