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BioAssay: AID 651749

qHTS assay for small molecule activators of the nuclear factor-kappa B (NF-kB) signaling pathway

Transcription factors and their stimulated target gene expression play pivotal roles across many cellular processes including responses to environmental stimuli. Proper regulation of associated pathways is critical to maintain cell health and function. Constitutive activation of transcription factor pathways has been implicated in tumors as it may block cellular functions such as apoptosis and lead to invasion and metastasis. ..more
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 Tested Compounds
 Tested Compounds
All(2329)
 
 
Active(8)
 
 
Inactive(2168)
 
 
Inconclusive(198)
 
 
 Tested Substances
 Tested Substances
All(2864)
 
 
Active(10)
 
 
Inactive(2634)
 
 
Inconclusive(220)
 
 
 Related BioAssays
 Related BioAssays
AID: 651749
Data Source: NCGC (NFkB916)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-05

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 8
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:

Transcription factors and their stimulated target gene expression play pivotal roles across many cellular processes including responses to environmental stimuli. Proper regulation of associated pathways is critical to maintain cell health and function. Constitutive activation of transcription factor pathways has been implicated in tumors as it may block cellular functions such as apoptosis and lead to invasion and metastasis.
One important transcription factor that stimulates gene expression in response to cytokines, UV radiation, and other stimuli is nuclear factor-kappa B (NF-kB). To measure activation of the NF-kB pathway, the NF-kB-bla ME-180 cell line stably expressed beta-lactamase reporter gene under the regulation of a NF-kB response element was obtained from Invitrogen. This assay allows measurement of the NF-kB pathway activity induced by compounds in a ME-180 cell line. This system quantitates NF-kB activity using a FRET-based assay system where beta-lactamase expression disrupts FRET, allowing for fluorescent measurement of activity.
Protocol
Assay Protocol Summary:

For toxicology screening, 2,000 cells in 5 uL/well were dispensed into black, clear-bottom 1536-well plates using a Multidrop Combi (Thermo Scientific) dispenser. Following transfer of 23 nL compound or DMSO vehicle by a pin tool, the plates were incubated 16 hr at 37C and 5% CO2. 1 ul LiveBLAzer FRET substrate (Invitrogen) was then added to each plate and following 90 min incubation at room temperature, fluorescence intensity (405nm excitation, 460 and 530 nm emission) was quantified on an Envision plate reader (PerkinElmer).
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 activators are ranked higher than compounds that showed apparent inhibition.
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: ME-180
From ChEMBL:
Assay Type: Functional
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*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
3EfficacyMaximal 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 CommentAnnotation/notes on a particular compound's data or its analysis.String
5W460-Activity_ScoreActivity score.Integer
6W460-Curve_DescriptionA 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
7W460-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8W460-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9W460-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10W460-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11W460-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12W460-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13W460-Excluded_PointsWhich 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
14W460-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15W460-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
16W460-Activity at 0.00130 uM (0.00129885μM**)% Activity at given concentration.Float%
17W460-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
18W460-Activity at 0.00490 uM (0.00490421μM**)% Activity at given concentration.Float%
19W460-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
20W460-Activity at 0.035 uM (0.0348284μM**)% Activity at given concentration.Float%
21W460-Activity at 0.057 uM (0.0569194μM**)% Activity at given concentration.Float%
22W460-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
23W460-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
24W460-Activity at 0.636 uM (0.636272μM**)% Activity at given concentration.Float%
25W460-Activity at 0.961 uM (0.960961μM**)% Activity at given concentration.Float%
26W460-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
27W460-Activity at 4.858 uM (4.85773μM**)% Activity at given concentration.Float%
28W460-Activity at 8.490 uM (8.49035μM**)% Activity at given concentration.Float%
29W460-Activity at 25.15 uM (25.1452μM**)% Activity at given concentration.Float%
30W460-Activity at 36.32 uM (36.3191μM**)% Activity at given concentration.Float%
31W460-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
32W460-Activity at 182.0 uM (182.026μM**)% Activity at given concentration.Float%
33W460-Activity at 406.9 uM (406.925μM**)% Activity at given concentration.Float%
34W460-Activity at 910.0 uM (910.026μM**)% Activity at given concentration.Float%
35W460-Activity at 2034.8 uM (2034.8μM**)% Activity at given concentration.Float%
36W460-Activity at 4831.4 uM (4831.42μM**)% Activity at given concentration.Float%
37W460-Activity at 10804.6 uM (10804.6μM**)% Activity at given concentration.Float%
38W460-Activity at 24160.9 uM (24160.9μM**)% Activity at given concentration.Float%
39W460-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
40Ratio-Activity_ScoreActivity score.Integer
41Ratio-Curve_DescriptionA 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
42Ratio-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
43Ratio-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
44Ratio-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
45Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
46Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
47Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
48Ratio-Excluded_PointsWhich 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
49Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
50Ratio-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
51Ratio-Activity at 0.00130 uM (0.00129885μM**)% Activity at given concentration.Float%
52Ratio-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
53Ratio-Activity at 0.00490 uM (0.00490421μM**)% Activity at given concentration.Float%
54Ratio-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
55Ratio-Activity at 0.035 uM (0.0348284μM**)% Activity at given concentration.Float%
56Ratio-Activity at 0.057 uM (0.0569194μM**)% Activity at given concentration.Float%
57Ratio-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
58Ratio-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
59Ratio-Activity at 0.636 uM (0.636272μM**)% Activity at given concentration.Float%
60Ratio-Activity at 0.961 uM (0.960961μM**)% Activity at given concentration.Float%
61Ratio-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
62Ratio-Activity at 4.858 uM (4.85773μM**)% Activity at given concentration.Float%
63Ratio-Activity at 8.490 uM (8.49035μM**)% Activity at given concentration.Float%
64Ratio-Activity at 25.15 uM (25.1452μM**)% Activity at given concentration.Float%
65Ratio-Activity at 36.32 uM (36.3191μM**)% Activity at given concentration.Float%
66Ratio-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
67Ratio-Activity at 182.0 uM (182.026μM**)% Activity at given concentration.Float%
68Ratio-Activity at 406.9 uM (406.925μM**)% Activity at given concentration.Float%
69Ratio-Activity at 910.0 uM (910.026μM**)% Activity at given concentration.Float%
70Ratio-Activity at 2034.8 uM (2034.8μM**)% Activity at given concentration.Float%
71Ratio-Activity at 4831.4 uM (4831.42μM**)% Activity at given concentration.Float%
72Ratio-Activity at 10804.6 uM (10804.6μM**)% Activity at given concentration.Float%
73Ratio-Activity at 24160.9 uM (24160.9μM**)% Activity at given concentration.Float%
74Ratio-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
75W530-Activity_ScoreActivity score.Integer
76W530-Curve_DescriptionA 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
77W530-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
78W530-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
79W530-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
80W530-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
81W530-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
82W530-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
83W530-Excluded_PointsWhich 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
84W530-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
85W530-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
86W530-Activity at 0.00130 uM (0.00129885μM**)% Activity at given concentration.Float%
87W530-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
88W530-Activity at 0.00490 uM (0.00490421μM**)% Activity at given concentration.Float%
89W530-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
90W530-Activity at 0.035 uM (0.0348284μM**)% Activity at given concentration.Float%
91W530-Activity at 0.057 uM (0.0569194μM**)% Activity at given concentration.Float%
92W530-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
93W530-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
94W530-Activity at 0.636 uM (0.636272μM**)% Activity at given concentration.Float%
95W530-Activity at 0.961 uM (0.960961μM**)% Activity at given concentration.Float%
96W530-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
97W530-Activity at 4.858 uM (4.85773μM**)% Activity at given concentration.Float%
98W530-Activity at 8.490 uM (8.49035μM**)% Activity at given concentration.Float%
99W530-Activity at 25.15 uM (25.1452μM**)% Activity at given concentration.Float%
100W530-Activity at 36.32 uM (36.3191μM**)% Activity at given concentration.Float%
101W530-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
102W530-Activity at 182.0 uM (182.026μM**)% Activity at given concentration.Float%
103W530-Activity at 406.9 uM (406.925μM**)% Activity at given concentration.Float%
104W530-Activity at 910.0 uM (910.026μM**)% Activity at given concentration.Float%
105W530-Activity at 2034.8 uM (2034.8μM**)% Activity at given concentration.Float%
106W530-Activity at 4831.4 uM (4831.42μM**)% Activity at given concentration.Float%
107W530-Activity at 10804.6 uM (10804.6μM**)% Activity at given concentration.Float%
108W530-Activity at 24160.9 uM (24160.9μM**)% Activity at given concentration.Float%
109W530-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
110Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

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