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BioAssay: AID 540368

Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 1 (S1P1) agonist assay Set 3

Name: Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 1 (S1P1) agonist assay Set 3. ..more
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 Tested Compounds
 Tested Compounds
All(30)
 
 
Active(5)
 
 
Inactive(25)
 
 
 Tested Substances
 Tested Substances
All(30)
 
 
Active(5)
 
 
Inactive(25)
 
 
AID: 540368
Data Source: The Scripps Research Institute Molecular Screening Center (S1P1_AG_BLA_384_3XEC50_SET 3)
BioAssay Type: Panel, Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-08-19
Hold-until Date: 2012-03-01
Modify Date: 2012-03-01

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 5
Related Experiments
Show more
AIDNameTypeProbeComment
373S1P3 Agonist Primary HTS and Confirmation AssaysScreening depositor-specified cross reference: Primary screen (S1P3 agonists in singlicate)
439S1P3 Agonist Dose-Response Potency AssayConfirmatory depositor-specified cross reference: Dose response (S1P3 agonists in triplicate)
1192Dose Response Cell-Based Assay for Agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Purchased AnaloguesConfirmatory depositor-specified cross reference: Dose response (S1P3 agonists in triplicate)
540309Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 3 (S1P3)Summary5 depositor-specified cross reference: Summary (S1P3 agonists in triplicate)
540344Late-stage assay provider results from the probe development effort to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): luminescence-based cell-based dose response counterscreen assay to determine cytotoxicity of agonist compoundsConfirmatory same project related to Summary assay
540349Late-stage fluorescence-based cell-based dose response assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Synthesized compoundsConfirmatory same project related to Summary assay
540351Late-stage counterscreen panel assay for S1P3 agonists: Ricerca HitProfilingScreen + CYP450Other same project related to Summary assay
540366Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 4 (S1P4) agonist assayConfirmatory same project related to Summary assay
540367Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 2 (S1P2) agonist assayConfirmatory same project related to Summary assay
540369Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 5 (S1P5) agonist assayConfirmatory same project related to Summary assay
588327Late-stage assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): radiometric-based cell-based dose response S1P agonist competition binding assayConfirmatory same project related to Summary assay
Description:
Source (MLSCN Center Name): The Scripps Research Institute Molecular Screening Center
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Michael Oldstone, TSRI
Network: Molecular Library Screening Center Network (MLSCN)
Grant Proposal Number: U01 AI074564
Grant Proposal PI: Michael Oldstone, TSRI

External Assay ID: S1P1_AG_BLA_384_3XEC50_SET 3

Name: Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 1 (S1P1) agonist assay Set 3.

Description:

Sphingosine 1-phosphate (S1P) is a lysophospholipid signaling molecule that regulates important biological functions in both intracellular (1) and extracellular compartments (2), including a wide variety of physiological responses such as heart rate (3-4), coronary artery caliber, endothelial integrity, and lymphocyte recirculation (4-7). These responses are mediated through high-affinity interactions with five members of the endothelial differentiation gene (EDG) family of plasma membrane-localized G-protein-coupled receptors (GPCRs), the sphingosine lipid receptors, S1P1-5 (8-10). S1P3 receptor couples promiscuously to Gi, Gq, and G12/13 proteins (11-13). Its expression is widespread (14-16). The S1P3 knockout mouse is phenotypically normal (14). Most S1P-mediated responses on endothelial cells occur via the S1P1 receptor alone or in combination with the S1P3 receptor. Bradycardia and hypertension are clearly associated with S1P3 activation and its expression patterns in cardiac tissue (3, 17). The use of the S1P1-selective agonist SEW2871 together with S1P3-deletant mice showed that activation of S1P3 regulates sinus rhythm, whereas activation of S1P1 plays no discernable role in the process (4). S1P3 on dendritic cells has been identified as a major exacerbating factor for mortality during sepsis by playing a role in the critical linkage of inflammation and coagulation pathways downstream of the thrombin cascade (18). A potent and selective S1P3 agonist would be useful in dissecting the complexities of S1P-mediated physiological processes in which S1P3 is involved, including bradycardia and hypertension.

References:

1. Goetzl, E. J., Wang, W., McGiffert, C., Liao, J. J., and Huang, M. C. (2007) Sphingosine 1-phosphate as an intracellular messenger and extracellular mediator in immunity, Acta Paediatr Suppl 96, 49-52
2. Spiegel, S., and Milstien, S. (2003) Sphingosine-1-phosphate: an enigmatic signalling lipid, Nat Rev Mol Cell Biol 4, 397-407.
3. Forrest, M., Sun, S. Y., Hajdu, R., Bergstrom, J., Card, D., Doherty, G., Hale, J., Keohane, C., Meyers, C., Milligan, J., Mills, S., Nomura, N., Rosen, H., Rosenbach, M., Shei, G. J., Singer, II, Tian, M., West, S., White, V., Xie, J., Proia, R. L., and Mandala, S. (2004) Immune cell regulation and cardiovascular effects of sphingosine 1-phosphate receptor agonists in rodents are mediated via distinct receptor subtypes, J Pharmacol Exp Ther 309, 758-768.
4. Sanna, M. G., Liao, J., Jo, E., Alfonso, C., Ahn, M. Y., Peterson, M. S., Webb, B., Lefebvre, S., Chun, J., Gray, N., and Rosen, H. (2004) Sphingosine 1-phosphate (S1P) receptor subtypes S1P1 and S1P3, respectively, regulate lymphocyte recirculation and heart rate, J Biol Chem 279, 13839-13848.
5. Alfonso, C., McHeyzer-Williams, M. G., and Rosen, H. (2006) CD69 down-modulation and inhibition of thymic egress by short- and long-term selective chemical agonism of sphingosine 1-phosphate receptors, Eur J Immunol 36, 149-159.
6. Jo, E., Sanna, M. G., Gonzalez-Cabrera, P. J., Thangada, S., Tigyi, G., Osborne, D. A., Hla, T., Parrill, A. L., and Rosen, H. (2005) S1P1-selective in vivo-active agonists from high-throughput screening: off-the-shelf chemical probes of receptor interactions, signaling, and fate, Chem Biol 12, 703-715.
7. Wei, S. H., Rosen, H., Matheu, M. P., Sanna, M. G., Wang, S. K., Jo, E., Wong, C. H., Parker, I., and Cahalan, M. D. (2005) Sphingosine 1-phosphate type 1 receptor agonism inhibits transendothelial migration of medullary T cells to lymphatic sinuses, Nat Immunol 6, 1228-1235.
8. Hla, T. (2003) Signaling and biological actions of sphingosine 1-phosphate, Pharmacol Res 47, 401-407.
9. Mandala, S., Hajdu, R., Bergstrom, J., Quackenbush, E., Xie, J., Milligan, J., Thornton, R., Shei, G. J., Card, D., Keohane, C., Rosenbach, M., Hale, J., Lynch, C. L., Rupprecht, K., Parsons, W., and Rosen, H. (2002) Alteration of lymphocyte trafficking by sphingosine-1-phosphate receptor agonists, Science 296, 346-349.
10. Sanchez, T., and Hla, T. (2004) Structural and functional characteristics of S1P receptors, J Cell Biochem 92, 913-922.
11. Kon, J., Sato, K., Watanabe, T., Tomura, H., Kuwabara, A., Kimura, T., Tamama, K., Ishizuka, T., Murata, N., Kanda, T., Kobayashi, I., Ohta, H., Ui, M., and Okajima, F. (1999) Comparison of intrinsic activities of the putative sphingosine 1-phosphate receptor subtypes to regulate several signaling pathways in their cDNA-transfected Chinese hamster ovary cells, J Biol Chem 274, 23940-23947.
12. Okamoto, H., Takuwa, N., Yatomi, Y., Gonda, K., Shigematsu, H., and Takuwa, Y. (1999) EDG3 is a functional receptor specific for sphingosine 1-phosphate and sphingosylphosphorylcholine with signaling characteristics distinct from EDG1 and AGR16, Biochem Biophys Res Commun 260, 203-208.
13. Windh, R. T., Lee, M. J., Hla, T., An, S., Barr, A. J., and Manning, D. R. (1999) Differential coupling of the sphingosine 1-phosphate receptors Edg-1, Edg-3, and H218/Edg-5 to the G(i), G(q), and G(12) families of heterotrimeric G proteins, J Biol Chem 274, 27351-27358.
14. Ishii, I., Friedman, B., Ye, X., Kawamura, S., McGiffert, C., Contos, J. J., Kingsbury, M. A., Zhang, G., Brown, J. H., and Chun, J. (2001) Selective loss of sphingosine 1-phosphate signaling with no obvious phenotypic abnormality in mice lacking its G protein-coupled receptor, LP(B3)/EDG-3, J Biol Chem 276, 33697-33704.
15. Zhang, G., Contos, J. J., Weiner, J. A., Fukushima, N., and Chun, J. (1999) Comparative analysis of three murine G-protein coupled receptors activated by sphingosine-1-phosphate, Gene 227, 89-99.
16. Yamaguchi, F., Tokuda, M., Hatase, O., and Brenner, S. (1996) Molecular cloning of the novel human G protein-coupled receptor (GPCR) gene mapped on chromosome 9, Biochem Biophys Res Commun 227, 608-614.
17. Murakami, A., Takasugi, H., Ohnuma, S., Koide, Y., Sakurai, A., Takeda, S., Hasegawa, T., Sasamori, J., Konno, T., Hayashi, K., Watanabe, Y., Mori, K., Sato, Y., Takahashi, A., Mochizuki, N., and Takakura, N. (2010) Sphingosine 1-phosphate (S1P) regulates vascular contraction via S1P3 receptor: investigation based on a new S1P3 receptor antagonist, Mol Pharmacol 77, 704-713.
18. Niessen, F., Schaffner, F., Furlan-Freguia, C., Pawlinski, R., Bhattacharjee, G., Chun, J., Derian, C. K., Andrade-Gordon, P., Rosen, H., and Ruf, W. (2008) Dendritic cell PAR1-S1P3 signalling couples coagulation and inflammation, Nature 452, 654-658.

Keywords:

Sphingosine Receptor, Sphingosine-1-phosphate receptor 3, S1P3, endothelial differentiation sphingolipid G-protein-coupled receptor 3, EDG3, Sphingosine-1-phosphate receptor 1, S1P1, EDG1, agonist, activator, GPCR, 384, Tango, FRET, GAL4-VP16, beta-arrestin, beta-lactamase, BLA, reporter gene, fluorescence, primary, late stage, late stage AID, bradycardia, hypertension, powders, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Library Screening Center Network, MLSCN
Panel Information
Experiments
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1Experiment 1624sphingosine 1-phosphate receptor 1 [Homo sapiens] [gi:13027636]
Taxonomy id: 9606
Gene id: 1901
2Experiment 2819sphingosine 1-phosphate receptor 1 [Homo sapiens] [gi:13027636]
Taxonomy id: 9606
Gene id: 1901
3Experiment 3821sphingosine 1-phosphate receptor 1 [Homo sapiens] [gi:13027636]
Taxonomy id: 9606
Gene id: 1901

§ Panel component ID.
Protocol
Assay Overview:
The purpose of this assay is to determine whether powder samples of compounds identified as active in the assay "Late-stage fluorescence dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Synthesized compounds" (AID 540349) were nonselective agonists as assayed by activation of S1P1. This assay uses Tango S1P1-bla U2OS cells which express the human Endothelial Differentiation Gene 1 (EDG1; S1P1) linked to a GAL4-VP16 transcription factor via a TEV protease site. The cells also express a beta-arrestin/TEV protease fusion protein and a beta-lactamase (BLA) reporter gene under the control of a UAS response element. Stimulation of the S1P1 receptor by agonist causes migration of the fusion protein to the GPCR, and through proteolysis liberates GAL4-VP16 from the receptor. The liberated VP16-GAL4 migrates to the nucleus, where it induces transcription of the BLA gene. BLA expression is monitored by measuring fluorescence resonance energy transfer (FRET) of a cleavable, fluorogenic, cell-permeable BLA substrate. As designed, test compounds that act as S1P1 agonists will activate S1P1 and increase well FRET. Compounds were tested in triplicate using a 10-point, 1:3 dilution series starting at a nominal concentration of 10 uM in Experiment 1, and 50 uM in Experiments 2 and 3.
Protocol Summary:
U2OS cells were cultured in T-175 sq cm flasks at 37 C and 95% relative humidity (RH). The growth media consisted of McCoy's 5A Medium supplemented with 10% v/v dialyzed fetal bovine serum, 0.1 mM NEAA, 25 mM HEPES (pH 7.3), 1 mM sodium pyruvate, 100 U/mL penicillin-streptomycin-neomycin, 200 ug/mL Zeocin, 50 ug/mL Hygromycin, and 100 ug/mL Geneticin.
Prior to the start of the assay, cells were suspended at a concentration of 275,000/mL in Assay Medium (Freestyle Expression Medium without supplements). The assay was started by dispensing 10 uL of cell suspension to each well of a 384 well plate (10,000 cells/well), followed by overnight incubation at 37 C in 5% CO2 and 95% RH. The next day, 50 nL of test compound in DMSO (0.5 % final DMSO concentration), DMSO alone, or S1P (40 nM final nominal EC80 concentration) prepared in 2% BSA was added to the appropriate wells. Plates were then incubated at 37 C in 5% CO2 for 4 hours. After the incubation, 2.2 uL/well of the LiveBLAzer FRET substrate mixture, prepared according to the manufacturer's protocol and containing 10 mM Probenicid, was added to all wells. After 2 hours of incubation at room temperature in the dark, plates were read on the EnVision plate reader (PerkinElmer Lifesciences, Turku, Finland) at an excitation wavelength of 405 nm and emission wavelengths of 460 nm and 535 nm.
Prior to normalization, data were corrected by subtracting "background" for both emission channels (ie, fluorescence values from cell-free wells containing media and substrate only). To normalize assay data, these corrected values were used to calculate a ratio for each well, according to the following mathematical expression:
Ratio = I460 nm / I535 nm
Where:
I represents the measured fluorescence emission intensity at the enumerated wavelength.
The percent activation for each compound was calculated using well fluorescence as follows:
%_Activation = 100 * ( 1 - ( ( Test_Compound - Median_High_Control ) / ( Median_Low_Control - Median_High_Control ) ) )
Where:
Test_Compound is defined as wells containing test compound and S1P
Low_Control is defined as wells containing DMSO
High_Control is defined as wells containing 5 uM S1P
Percent activation was plotted against the log of the compound concentration. A three parameter equation describing a sigmoidal dose-response curve was then fitted using GraphPad Prism (GraphPad Software Inc) normalized from 0 to 100 for each assay. The software-generated EC50 values were reported. In cases where the highest concentration tested (i.e. 10 uM in Experiment 1 and 50 uM in Experiments 2 and 2) did not result in greater than 50% activation, the EC50 was determined manually as greater than the highest concentration tested. In cases where the software was not able to generate a curve, "Not Converged" is reported for the statistical values.
PubChem Activity Outcome and Score:
The following applies to each panel in this assay:
Compounds with an EC50 greater than 10 uM were considered inactive. Compounds with an EC50 equal to or less than 10 uM were considered active
Activity score was then ranked by the potency of the compounds with fitted curves, with the most potent compounds assigned the highest activity scores.
Experiment 1 Score: The PubChem Activity Score range for active compounds is 100-1, and for inactive compounds 0-0.
Experiment 2 Score: The PubChem Activity Score range for active compounds is 100-84, and for inactive compounds 41-0.
Experiment 3 Score: The PubChem Activity Score range for active compounds is 100-87, and for inactive compounds 82-0.
Overall Outcome and Score:
Compounds that were active in all experiments were considered active, otherwise they were considered inactive.
The overall score is 0 if the compound was inactive, otherwise the score is taken as the fraction of panels where the compound is active, multiplied by 100.
The PubChem Activity Score is assigned a value of 100 for active compounds, and 0 for inactive compounds.
The PubChem Activity Score range for active compounds is 100-100, and for inactive compounds 0-0.
List of Reagents:
Tango EDG1-bla U2OS cells (Invitrogen, part K1520)
GeneBLAzer FRET B/G Loading Kit (CCF4-AM) (Invitrogen, part K1025)
Probenecid (Sigma, part P8761)
Freestyle Expression Medium (Assay media; Invitrogen, part 12338-018)
McCoy's 5A Medium (modified) (1X) (Invitrogen, 16600-082)
Fetal Bovine Serum, dialyzed (Invitrogen, part 26400-036)
NEAA (Invitrogen, part 1114-050)
Penicillin-Streptomycin-Neomycin antibiotic mix (Invitrogen, part 15140-122)
Sodium Pyruvate (Invitrogen, part 11360-070)
PBS without calcium or magnesium (Invitrogen, part 14190-136)
HEPES (Invitrogen, part 15630-080)
Trypsin/EDTA (Invitrogen, part 25300-054)
S1P (Avanti Polar Lipids, part 860492P)
Fatty Acid Free BSA (Calbiochem, part NC9734015)
Zeocin (Invitrogen, part R250-01)
Hygromycin (Invitrogen, part 10687-010)
Geneticin (Invitrogen, part 10131-027)
384-well plates (Greiner, part 788092)
T175 tissue culture flasks (Corning, part 431080)
Comment
In this assay, S1P had a 50% effective concentration (EC50) of approximately 50 nM. Possible artifacts of this assay can include, but are not limited to: dust or lint located in or on wells of the microtiter plate, compounds that modulate beta-arrestin or BLA activity, and compounds that quench or emit fluorescence.
Categorized Comment - additional comments and annotations
From BioAssay Depositor:
Assay: CurveFit [1]: Equation: = 100 / ( 1 + 10^( ( [Log EC50] - Log( [Concentration] * 10^-6 ) * [Hill Slope] ) )
Assay: Dictionary: Version: 0.1
From PubChem:
Assay Format: Cell-based
Assay Cell Type: U-2 OS
Result Definitions
Show more
TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Outcome [Exp 1]The outcome of the experiment, one of "active" or "inactive".1sphingosine 1-phosphate receptor 1 [Homo sapiens]Outcome
2Score [Exp 1]The BioAssay activity ranking score1Integer
3Qualifier [Exp 1]Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration1String
4EC50 [Exp 1]*The concentration at which 50 percent of the activity in the antagonist assay is observed; (EC50) shown in micromolar.1FloatμM
5Log EC50 [Exp 1]The Log of EC501Float
6Hill Slope [Exp 1]The Hill Slope1Float
7R squared [Exp 1]The value of R squared1Float
8Activation at 10 uM [1, Exp 1] (10μM**)Value of % Activation at 10 uM activator concentration [1, Exp 1]1Float%
9Activation at 10 uM [2, Exp 1] (10μM**)Value of % Activation at 10 uM activator concentration [2, Exp 1]1Float%
10Activation at 10 uM [3, Exp 1] (10μM**)Value of % Activation at 10 uM activator concentration [3, Exp 1]1Float%
11Activation at 10 uM [4, Exp 1] (10μM**)Value of % Activation at 10 uM activator concentration [4, Exp 1]1Float%
12Activation at 3.3 uM [1, Exp 1] (3.3μM**)Value of % Activation at 3.3 uM activator concentration [1, Exp 1]1Float%
13Activation at 3.3 uM [2, Exp 1] (3.3μM**)Value of % Activation at 3.3 uM activator concentration [2, Exp 1]1Float%
14Activation at 3.3 uM [3, Exp 1] (3.3μM**)Value of % Activation at 3.3 uM activator concentration [3, Exp 1]1Float%
15Activation at 3.3 uM [4, Exp 1] (3.3μM**)Value of % Activation at 3.3 uM activator concentration [4, Exp 1]1Float%
16Activation at 1.1 uM [1, Exp 1] (1.1μM**)Value of % Activation at 1.1 uM activator concentration [1, Exp 1]1Float%
17Activation at 1.1 uM [2, Exp 1] (1.1μM**)Value of % Activation at 1.1 uM activator concentration [2, Exp 1]1Float%
18Activation at 1.1 uM [3, Exp 1] (1.1μM**)Value of % Activation at 1.1 uM activator concentration [3, Exp 1]1Float%
19Activation at 1.1 uM [4, Exp 1] (1.1μM**)Value of % Activation at 1.1 uM activator concentration [4, Exp 1]1Float%
20Activation at 0.367 uM [1, Exp 1] (0.367μM**)Value of % Activation at 0.367 uM activator concentration [1, Exp 1]1Float%
21Activation at 0.367 uM [2, Exp 1] (0.367μM**)Value of % Activation at 0.367 uM activator concentration [2, Exp 1]1Float%
22Activation at 0.367 uM [3, Exp 1] (0.367μM**)Value of % Activation at 0.367 uM activator concentration [3, Exp 1]1Float%
23Activation at 0.367 uM [4, Exp 1] (0.367μM**)Value of % Activation at 0.367 uM activator concentration [4, Exp 1]1Float%
24Activation at 0.122 uM [1, Exp 1] (0.122μM**)Value of % Activation at 0.122 uM activator concentration [1, Exp 1]1Float%
25Activation at 0.122 uM [2, Exp 1] (0.122μM**)Value of % Activation at 0.122 uM activator concentration [2, Exp 1]1Float%
26Activation at 0.122 uM [3, Exp 1] (0.122μM**)Value of % Activation at 0.122 uM activator concentration [3, Exp 1]1Float%
27Activation at 0.122 uM [4, Exp 1] (0.122μM**)Value of % Activation at 0.122 uM activator concentration [4, Exp 1]1Float%
28Activation at 0.0408 uM [1, Exp 1] (0.0408μM**)Value of % Activation at 0.0408 uM activator concentration [1, Exp 1]1Float%
29Activation at 0.0408 uM [2, Exp 1] (0.0408μM**)Value of % Activation at 0.0408 uM activator concentration [2, Exp 1]1Float%
30Activation at 0.0408 uM [3, Exp 1] (0.0408μM**)Value of % Activation at 0.0408 uM activator concentration [3, Exp 1]1Float%
31Activation at 0.0408 uM [4, Exp 1] (0.0408μM**)Value of % Activation at 0.0408 uM activator concentration [4, Exp 1]1Float%
32Activation at 0.0136 uM [1, Exp 1] (0.0136μM**)Value of % Activation at 0.0136 uM activator concentration [1, Exp 1]1Float%
33Activation at 0.0136 uM [2, Exp 1] (0.0136μM**)Value of % Activation at 0.0136 uM activator concentration [2, Exp 1]1Float%
34Activation at 0.0136 uM [3, Exp 1] (0.0136μM**)Value of % Activation at 0.0136 uM activator concentration [3, Exp 1]1Float%
35Activation at 0.0136 uM [4, Exp 1] (0.0136μM**)Value of % Activation at 0.0136 uM activator concentration [4, Exp 1]1Float%
36Activation at 0.0045 uM [1, Exp 1] (0.0045μM**)Value of % Activation at 0.0045 uM activator concentration [1, Exp 1]1Float%
37Activation at 0.0045 uM [2, Exp 1] (0.0045μM**)Value of % Activation at 0.0045 uM activator concentration [2, Exp 1]1Float%
38Activation at 0.0045 uM [3, Exp 1] (0.0045μM**)Value of % Activation at 0.0045 uM activator concentration [3, Exp 1]1Float%
39Activation at 0.0045 uM [4, Exp 1] (0.0045μM**)Value of % Activation at 0.0045 uM activator concentration [4, Exp 1]1Float%
40Activation at 0.0015 uM [1, Exp 1] (0.0015μM**)Value of % Activation at 0.0015 uM activator concentration [1, Exp 1]1Float%
41Activation at 0.0015 uM [2, Exp 1] (0.0015μM**)Value of % Activation at 0.0015 uM activator concentration [2, Exp 1]1Float%
42Activation at 0.0015 uM [3, Exp 1] (0.0015μM**)Value of % Activation at 0.0015 uM activator concentration [3, Exp 1]1Float%
43Activation at 0.0015 uM [4, Exp 1] (0.0015μM**)Value of % Activation at 0.0015 uM activator concentration [4, Exp 1]1Float%
44Activation at 0.000504 uM [1, Exp 1] (0.000504μM**)Value of % Activation at 0.000504 uM activator concentration [1, Exp 1]1Float%
45Activation at 0.000504 uM [2, Exp 1] (0.000504μM**)Value of % Activation at 0.000504 uM activator concentration [2, Exp 1]1Float%
46Activation at 0.000504 uM [3, Exp 1] (0.000504μM**)Value of % Activation at 0.000504 uM activator concentration [3, Exp 1]1Float%
47Activation at 0.000504 uM [4, Exp 1] (0.000504μM**)Value of % Activation at 0.000504 uM activator concentration [4, Exp 1]1Float%
48Outcome [Exp 2]The outcome of the experiment, one of "active" or "inactive".2sphingosine 1-phosphate receptor 1 [Homo sapiens]Outcome
49Score [Exp 2]The BioAssay activity ranking score2Integer
50Qualifier [Exp 2]Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration2String
51EC50 [Exp 2]*The concentration at which 50 percent of the activity in the antagonist assay is observed; (EC50) shown in micromolar.2FloatμM
52Log EC50 [Exp 2]The Log of EC502Float
53Hill Slope [Exp 2]The Hill Slope2Float
54R squared [Exp 2]The value of R squared2Float
55Activation at 50 uM [1, Exp 2] (50μM**)Value of % Activation at 50 uM activator concentration [1, Exp 2]2Float%
56Activation at 50 uM [2, Exp 2] (50μM**)Value of % Activation at 50 uM activator concentration [2, Exp 2]2Float%
57Activation at 50 uM [3, Exp 2] (50μM**)Value of % Activation at 50 uM activator concentration [3, Exp 2]2Float%
58Activation at 16.7 uM [1, Exp 2] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [1, Exp 2]2Float%
59Activation at 16.7 uM [2, Exp 2] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [2, Exp 2]2Float%
60Activation at 16.7 uM [3, Exp 2] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [3, Exp 2]2Float%
61Activation at 5.6 uM [1, Exp 2] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [1, Exp 2]2Float%
62Activation at 5.6 uM [2, Exp 2] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [2, Exp 2]2Float%
63Activation at 5.6 uM [3, Exp 2] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [3, Exp 2]2Float%
64Activation at 1.9 uM [1, Exp 2] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [1, Exp 2]2Float%
65Activation at 1.9 uM [2, Exp 2] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [2, Exp 2]2Float%
66Activation at 1.9 uM [3, Exp 2] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [3, Exp 2]2Float%
67Activation at 0.617 uM [1, Exp 2] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [1, Exp 2]2Float%
68Activation at 0.617 uM [2, Exp 2] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [2, Exp 2]2Float%
69Activation at 0.617 uM [3, Exp 2] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [3, Exp 2]2Float%
70Activation at 0.206 uM [1, Exp 2] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [1, Exp 2]2Float%
71Activation at 0.206 uM [2, Exp 2] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [2, Exp 2]2Float%
72Activation at 0.206 uM [3, Exp 2] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [3, Exp 2]2Float%
73Activation at 0.0686 uM [1, Exp 2] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [1, Exp 2]2Float%
74Activation at 0.0686 uM [2, Exp 2] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [2, Exp 2]2Float%
75Activation at 0.0686 uM [3, Exp 2] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [3, Exp 2]2Float%
76Activation at 0.0229 uM [1, Exp 2] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [1, Exp 2]2Float%
77Activation at 0.0229 uM [2, Exp 2] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [2, Exp 2]2Float%
78Activation at 0.0229 uM [3, Exp 2] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [3, Exp 2]2Float%
79Activation at 0.0076 uM [1, Exp 2] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [1, Exp 2]2Float%
80Activation at 0.0076 uM [2, Exp 2] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [2, Exp 2]2Float%
81Activation at 0.0076 uM [3, Exp 2] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [3, Exp 2]2Float%
82Activation at 0.0025 uM [1, Exp 2] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [1, Exp 2]2Float%
83Activation at 0.0025 uM [2, Exp 2] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [2, Exp 2]2Float%
84Activation at 0.0025 uM [3, Exp 2] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [3, Exp 2]2Float%
85Outcome [Exp 3]The outcome of the experiment, one of "active" or "inactive".3sphingosine 1-phosphate receptor 1 [Homo sapiens]Outcome
86Score [Exp 3]The BioAssay activity ranking score3Integer
87Qualifier [Exp 3]Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration3String
88EC50 [Exp 3]*The concentration at which 50 percent of the activity in the antagonist assay is observed; (EC50) shown in micromolar.3FloatμM
89Log EC50 [Exp 3]The Log of EC503Float
90Hill Slope [Exp 3]The Hill Slope3Float
91R squared [Exp 3]The value of R squared3Float
92Activation at 50 uM [1, Exp 3] (50μM**)Value of % Activation at 50 uM activator concentration [1, Exp 3]3Float%
93Activation at 50 uM [2, Exp 3] (50μM**)Value of % Activation at 50 uM activator concentration [2, Exp 3]3Float%
94Activation at 50 uM [3, Exp 3] (50μM**)Value of % Activation at 50 uM activator concentration [3, Exp 3]3Float%
95Activation at 16.7 uM [1, Exp 3] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [1, Exp 3]3Float%
96Activation at 16.7 uM [2, Exp 3] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [2, Exp 3]3Float%
97Activation at 16.7 uM [3, Exp 3] (16.7μM**)Value of % Activation at 16.7 uM activator concentration [3, Exp 3]3Float%
98Activation at 5.6 uM [1, Exp 3] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [1, Exp 3]3Float%
99Activation at 5.6 uM [2, Exp 3] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [2, Exp 3]3Float%
100Activation at 5.6 uM [3, Exp 3] (5.6μM**)Value of % Activation at 5.6 uM activator concentration [3, Exp 3]3Float%
101Activation at 1.9 uM [1, Exp 3] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [1, Exp 3]3Float%
102Activation at 1.9 uM [2, Exp 3] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [2, Exp 3]3Float%
103Activation at 1.9 uM [3, Exp 3] (1.9μM**)Value of % Activation at 1.9 uM activator concentration [3, Exp 3]3Float%
104Activation at 0.617 uM [1, Exp 3] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [1, Exp 3]3Float%
105Activation at 0.617 uM [2, Exp 3] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [2, Exp 3]3Float%
106Activation at 0.617 uM [3, Exp 3] (0.617μM**)Value of % Activation at 0.617 uM activator concentration [3, Exp 3]3Float%
107Activation at 0.206 uM [1, Exp 3] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [1, Exp 3]3Float%
108Activation at 0.206 uM [2, Exp 3] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [2, Exp 3]3Float%
109Activation at 0.206 uM [3, Exp 3] (0.206μM**)Value of % Activation at 0.206 uM activator concentration [3, Exp 3]3Float%
110Activation at 0.0686 uM [1, Exp 3] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [1, Exp 3]3Float%
111Activation at 0.0686 uM [2, Exp 3] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [2, Exp 3]3Float%
112Activation at 0.0686 uM [3, Exp 3] (0.0686μM**)Value of % Activation at 0.0686 uM activator concentration [3, Exp 3]3Float%
113Activation at 0.0229 uM [1, Exp 3] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [1, Exp 3]3Float%
114Activation at 0.0229 uM [2, Exp 3] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [2, Exp 3]3Float%
115Activation at 0.0229 uM [3, Exp 3] (0.0229μM**)Value of % Activation at 0.0229 uM activator concentration [3, Exp 3]3Float%
116Activation at 0.0076 uM [1, Exp 3] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [1, Exp 3]3Float%
117Activation at 0.0076 uM [2, Exp 3] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [2, Exp 3]3Float%
118Activation at 0.0076 uM [3, Exp 3] (0.0076μM**)Value of % Activation at 0.0076 uM activator concentration [3, Exp 3]3Float%
119Activation at 0.0025 uM [1, Exp 3] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [1, Exp 3]3Float%
120Activation at 0.0025 uM [2, Exp 3] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [2, Exp 3]3Float%
121Activation at 0.0025 uM [3, Exp 3] (0.0025μM**)Value of % Activation at 0.0025 uM activator concentration [3, Exp 3]3Float%

* Activity Concentration. ** Test Concentration. § Panel component ID.
Additional Information
Grant Number: U01 AI074564

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