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BioAssay: AID 504924

Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 3.7 nM

Name: Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 3.7 nM. ..more
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 Tested Compounds
 Tested Compounds
All(1)
 
 
Inactive(1)
 
 
 Tested Substances
 Tested Substances
All(1)
 
 
Inactive(1)
 
 
AID: 504924
Data Source: The Scripps Research Institute Molecular Screening Center (S1P4_AG_BLA_384_3XEC50_3.7nM Antagonist)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-07-06
Hold-until Date: 2012-03-01
Modify Date: 2012-03-01

Data Table ( Complete ):           View All Data
Target
Tested Compound:
Related Experiments
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AIDNameTypeProbeComment
1509Primary Cell-Based Assay to Identify Agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4)Screening depositor-specified cross reference: Primary screen (S1P4 agonists in singlicate)
1523Confirmation cell-based high throughput assay for agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4)Screening depositor-specified cross reference: Confirmation screen (S1P4 agonists in triplicate)
1563Counterscreen assay for S1P4 agonists: Cell-based high throughput screening assay to identify agonists of the Sphingosine 1-Phosphate Receptor 1 (S1P1)Screening depositor-specified cross reference: Counterscreen (S1P1 agonists in triplicate)
1686Fluorescence dose response cell-based high throughput screening assay for agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4)Confirmatory depositor-specified cross reference: Dose response (S1P4 agonists in triplicate)
1701Fluorescence-based counterscreen assay for S1P4 agonists: Cell-based dose response high throughput screening assay to identify agonists of the Sphingosine 1-Phosphate Receptor 1 (S1P1)Confirmatory depositor-specified cross reference: Dose response counterscreen (S1P1 agonists in triplicate)
1801Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 4 (S1P4)Summary2 depositor-specified cross reference: Summary (S1P4 agonists)
463107Late-stage fluorescence dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Synthesized compoundsConfirmatory depositor-specified cross reference: Late stage dose response (S1P4 agonists in triplicate)
463118Late-stage fluorescence-based dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 1 (S1P1) counterscreen assayConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (S1P1 agonists in triplicate)
463119Late-stage assay provider results from the probe development effort to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): luminescence-based cell-based dose response assay to determine cytotoxicity of agonist compoundsConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (Cytotoxicity in quadruplicate)
463122Late-stage fluorescence-based dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 2 (S1P2) counterscreen assayConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (S1P2 agonists in triplicate)
463123Late-stage fluorescence-based dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 3 (S1P3) counterscreen assayConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (S1P3 agonists in triplicate)
463129Late-stage fluorescence-based dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 5 (S1P5) counterscreen assayConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (S1P5 agonists in triplicate)
463225Late-stage fluorescence dose response cell-based counterscreening assay for agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonistConfirmatory depositor-specified cross reference: Late stage dose response counterscreen (inhibition by S1P4-selective antagonist)
504400Late-stage counterscreen panel assay for S1P4 agonists: Ricerca HitProfilingScreen + CYP450Other depositor-specified cross reference: Late-stage counterscreen panel assay (Ricerca hit profiling CYP450)
504460Late-stage fluorescence dose response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): purchased compounds EC50Confirmatory depositor-specified cross reference: Late-stage dose response (S1P4 agonists in triplicate)
540332Late-stage counterscreen panel assay for S1P4 agonists: Ricerca HitProfilingScreen + CYP450: Set 2Other depositor-specified cross reference
504867Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 5 (S1P5) agonist assay Set 2Confirmatory same project related to Summary assay
504869Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 5 (S1P5) antagonist assay Set 2Confirmatory same project related to Summary assay
504870Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 1 (S1P1) agonist assay Set 2Confirmatory same project related to Summary assay
504871Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist controlConfirmatory same project related to Summary assay
504872Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 3 (S1P3) antagonist assay Set 2Confirmatory same project related to Summary assay
504873Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 2 (S1P2) agonist assay Set 2Confirmatory same project related to Summary assay
504875Late-stage assay provider results from the probe development effort to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): luminescence-based cell-based dose response assay to determine cytotoxicity of agonist compounds Set 2Confirmatory same project related to Summary assay
504876Late-stage fluorescence dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 4 (S1P4) antagonist assay Set 2Confirmatory same project related to Summary assay
504877Late-stage fluorescence dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Synthesized compounds set 2Other same project related to Summary assay
504879Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 3 (S1P3) agonist assay Set 2Confirmatory same project related to Summary assay
504880Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 1 (S1P1) antagonist assayConfirmatory same project related to Summary assay
504881Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Sphingosine 1-Phosphate Receptor 2 (S1P2) antagonist assay Set 2Confirmatory same project related to Summary assay
504917Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 1.2 nMConfirmatory same project related to Summary assay
504918Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 0.4 nMConfirmatory same project related to Summary assay
504919Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 11 nMConfirmatory same project related to Summary assay
504921Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 33 nMConfirmatory same project related to Summary assay
504923Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 100 nMConfirmatory same project related to Summary assay
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRISMC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Michael Oldstone, TSRI
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: U01 AI074564 Fast Track
Grant Proposal PI: Michael Oldstone, TSRI
External Assay Id: S1P4_AG_BLA_384_3XEC50_3.7nM Antagonist

Name: Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): inhibition by S1P4-selective antagonist 3.7 nM.

Description:

Pandemic influenza represents a significant public health threat, due in part to immune cell-mediated lung tissue damage induced during viral infection. Sphingosine 1-phosphate (S1P) is a bioactive phospholipid released by activated blood platelets and serves to influence endothelial integrity, lung epithelial integrity (1), and lymphocyte recirculation (2-5) through five related high affinity G-protein coupled receptors. Recently, modulation of S1P receptors locally in the lungs was shown to alter dendritic cell activation and accumulation in the mediastinal lymph nodes, resulting in blunted T cell responses and control of immunopathological features of influenza virus infection (6). Reports showing that S1P5 expression is very low in dendritic cells but that S1P4 is highly expressed (7), suggest that chemical activation of the S1P4 receptor subtype in the airways could be efficient at controlling the immunopathological response to viral infection. S1P4 is coupled to Gai and Gao G-proteins and activates ERK MAPK and PLC downstream pathways (8). Thus, the identification of compounds that act as selective S1P4 agonists will provide insight into S1P4 biology and may serve as useful tools to limit lung tissue injury resulting from influenza infection.

References:

1. Sanna, M.G., J. Liao, E. Jo, C. Alfonso, M.Y. Ahn, M.S. Peterson, B. Webb, S. Lefebvre, J. Chun, N. Gray, and H. Rosen, Sphingosine 1-phosphate (S1P) receptor subtypes S1P1 and S1P3, respectively, regulate lymphocyte recirculation and heart rate. J Biol Chem, 2004. 279(14): p. 13839-48.
2. Forrest, M., S.Y. Sun, R. Hajdu, J. Bergstrom, D. Card, G. Doherty, J. Hale, C. Keohane, C. Meyers, J. Milligan, S. Mills, N. Nomura, H. Rosen, M. Rosenbach, G.J. Shei, Singer, II, M. Tian, S. West, V. White, J. Xie, R.L. Proia, and S. Mandala, Immune cell regulation and cardiovascular effects of sphingosine 1-phosphate receptor agonists in rodents are mediated via distinct receptor subtypes. J Pharmacol Exp Ther, 2004. 309(2): p. 758-68.
3. Gon, Y., M.R. Wood, W.B. Kiosses, E. Jo, M.G. Sanna, J. Chun, and H. Rosen, S1P3 receptor-induced reorganization of epithelial tight junctions compromises lung barrier integrity and is potentiated by TNF. Proc Natl Acad Sci U S A, 2005. 102(26): p. 9270-5.
4. Wei, S.H., H. Rosen, M.P. Matheu, M.G. Sanna, S.K. Wang, E. Jo, C.H. Wong, I. Parker, and M.D. Cahalan, Sphingosine 1-phosphate type 1 receptor agonism inhibits transendothelial migration of medullary T cells to lymphatic sinuses. Nat Immunol, 2005. 6(12): p. 1228-35.
5. Alfonso, C., M.G. McHeyzer-Williams, and H. Rosen, CD69 down-modulation and inhibition of thymic egress by short- and long-term selective chemical agonism of sphingosine 1-phosphate receptors. Eur J Immunol, 2006. 36(1): p. 149-59.
6. Jo, E., M.G. Sanna, P.J. Gonzalez-Cabrera, S. Thangada, G. Tigyi, D.A. Osborne, T. Hla, A.L. Parrill, and H. Rosen, S1P1-selective in vivo-active agonists from high-throughput screening: off-the-shelf chemical probes of receptor interactions, signaling, and fate. Chem Biol, 2005. 12(6): p. 703-15.
7. Maeda, Y., Matsuyuki, H., Shimano, K., Kataoka, H., Sugahara, K., and Chiba, K., Migration of CD4 T cells and dendritic cells toward sphingosine 1-phosphate (S1P) is mediated by different receptor subtypes: S1P regulates the functions of murine mature dendritic cells via S1P receptor type 3. J Immunol, 2007. 178(6): p. 3437-46.
8. Toman, R.E. and S. Spiegel, Lysophospholipid receptors in the nervous system. Neurochem Res, 2002. 27(7-8): p. 619-27.

Keywords:

Sphingosine Receptor, Sphingosine-1-phosphate receptor 4, S1P4, EDG6, LPC1, agonist, activator, GPCR, 384, antagonist, Tango, FRET, GAL4-VP16, beta-arrestin, beta-lactamase, BLA, reporter gene, fluorescence, dose response, late stage, late stage AID, powders, Scripps, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Library Probe Production Centers Network, MLPCN
Protocol
Assay Overview:

The purpose of this assay is to determine whether a test compound with S1P4 agonist activity is inhibited by an S1P4-selective antagonist. This assay uses Tango S1P4-BLA U2OS cells which contain the human Endothelial Differentiation Gene 6 (EDG6; S1P4) 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 S1P4 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. The presence of an S1P4-selective antagonist compound in the assay inhibits S1P4 activation and migration of the fusion protein, thus preventing proteolysis of GAL4-VP16 and BLA transcription, leading to no increase in well FRET. In the presence of a test agonist compound, the S1P4 receptor will be stimulated, leading to an increase in well FRET. Compound was tested in triplicate using a 14-point, 1:3 dilution series starting at a nominal concentration of 3.3 uM.

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 1,000,000/mL in Assay Medium (Freestyle Expression Medium without supplements). The assay was started by dispensing 10 uL of cell suspension to each well, followed by overnight incubation at 37 C in 5% CO2 and 95% RH. The next day, 50 nL of test compound in DMSO was added to sample wells, and DMSO alone (0.5% final concentration) was added to the appropriate control wells. Next, S1P4-selective antagonist SID 117682935 was added to the appropriate wells to result in a final concentration of 3.7 nM. 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. The protocol summary and data for the control assay can be found in AID 504871.

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 -( ( Median_Test_Compound - Median_High_Control ) / ( Median_Low_Control - Median_High_Control ) )

Where:

Test_Compound is defined as wells containing test compound and S1P4-selective antagonist SID 117682935
Low_Control is defined as wells containing S1P4-selective antagonist SID 117682935
High_Control is defined as wells containing DMSO only.

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. 3.3 uM) did not result in greater than 50% activation, the EC50 was determined manually as greater than 3.3 uM.

PubChem Activity Outcome and Score:

The result of assays that resulted in EC50 greater than 3.3 uM were considered inactive. The result of assays that resulted in an EC50 equal to or less than 3.3 uM were considered active.

Any compound with a percent activity value < 50% at all test concentrations was assigned an activity score of zero. Any compound with a percent activity value >= 50% at any test concentration was assigned an activity score greater than zero.

Activity score was then ranked by the potency of the compounds with fitted curves, with the most potent compounds assigned the highest activity scores.

The PubChem Activity Score range for inactive compounds is 0-0. There are no active compounds.

List of Reagents:

Tango EDG6-bla U2OS cells (Invitrogen, part K1622)
S1P4-selective antagonist SID 117682935 (provided by SRIMSC)
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)
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
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^( ( [LogEC50] - Log( [Concentration] * 10^-6 ) * [Hill Slope] ) )
Assay: Dictionary: Version: 0.1
From PubChem:
Assay Format: Cell-based
Assay Cell Type: U-2 OS
From ChEMBL:
Assay Format: Cell-based
Assay Type: Functional
Result Definitions
Show more
TIDNameDescriptionAnnotationHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1Antagonist SIDAntagonist SIDSIDString
2Antagonist ConcentrationThe concentration of the S1P4-selective antagonist SID 117682935 in the assay, expressed in micromolar.FloatμM
3QualifierActivity 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 concentration.String
4EC50*The concentration at which 50 percent of activity in the agonist assay is observed; (EC50) shown in micromolar.FloatμM
5LogEC50Value of Log EC50Float
6Hill SlopeValue of Hill SlopeFloat
7R squaredValue of R squaredFloat
8Activation at 3.3 uM Ag [1] (3.3μM**)Value of % activation at 3.3 uM Agonist concentration; replicate [1]Float%
9Activation at 3.3 uM Ag [2] (3.3μM**)Value of % activation at 3.3 uM Agonist concentration; replicate [2]Float%
10Activation at 3.3 uM Ag [3] (3.3μM**)Value of % activation at 3.3 uM Agonist concentration; replicate [3]Float%
11Activation at 1.1 uM Ag [1] (1.1μM**)Value of % activation at 1.1 uM Agonist concentration; replicate [1]Float%
12Activation at 1.1 uM Ag [2] (1.1μM**)Value of % activation at 1.1 uM Agonist concentration; replicate [2]Float%
13Activation at 1.1 uM Ag [3] (1.1μM**)Value of % activation at 1.1 uM Agonist concentration; replicate [3]Float%
14Activation at 0.372 uM Ag [1] (0.372μM**)Value of % activation at 0.372 uM Agonist concentration; replicate [1]Float%
15Activation at 0.372 uM Ag [2] (0.372μM**)Value of % activation at 0.372 uM Agonist concentration; replicate [2]Float%
16Activation at 0.372 uM Ag [3] (0.372μM**)Value of % activation at 0.372 uM Agonist concentration; replicate [3]Float%
17Activation at 0.123 uM Ag [1] (0.123μM**)Value of % activation at 0.123 uM Agonist concentration; replicate [1]Float%
18Activation at 0.123 uM Ag [2] (0.123μM**)Value of % activation at 0.123 uM Agonist concentration; replicate [2]Float%
19Activation at 0.123 uM Ag [3] (0.123μM**)Value of % activation at 0.123 uM Agonist concentration; replicate [3]Float%
20Activation at 0.0407 uM Ag [1] (0.0407μM**)Value of % activation at 0.0407 uM Agonist concentration; replicate [1]Float%
21Activation at 0.0407 uM Ag [2] (0.0407μM**)Value of % activation at 0.0407 uM Agonist concentration; replicate [2]Float%
22Activation at 0.0407 uM Ag [3] (0.0407μM**)Value of % activation at 0.0407 uM Agonist concentration; replicate [3]Float%
23Activation at 0.0138 uM Ag [1] (0.0138μM**)Value of % activation at 0.0138 uM Agonist concentration; replicate [1]Float%
24Activation at 0.0138 uM Ag [2] (0.0138μM**)Value of % activation at 0.0138 uM Agonist concentration; replicate [2]Float%
25Activation at 0.0138 uM Ag [3] (0.0138μM**)Value of % activation at 0.0138 uM Agonist concentration; replicate [3]Float%
26Activation at 0.0046 uM Ag [1] (0.0046μM**)Value of % activation at 0.0046 uM Agonist concentration; replicate [1]Float%
27Activation at 0.0046 uM Ag [2] (0.0046μM**)Value of % activation at 0.0046 uM Agonist concentration; replicate [2]Float%
28Activation at 0.0046 uM Ag [3] (0.0046μM**)Value of % activation at 0.0046 uM Agonist concentration; replicate [3]Float%
29Activation at 0.0015 uM Ag [1] (0.0015μM**)Value of % activation at 0.0015 uM Agonist concentration; replicate [1]Float%
30Activation at 0.0015 uM Ag [2] (0.0015μM**)Value of % activation at 0.0015 uM Agonist concentration; replicate [2]Float%
31Activation at 0.0015 uM Ag [3] (0.0015μM**)Value of % activation at 0.0015 uM Agonist concentration; replicate [3]Float%
32Activation at 0.000513 uM Ag [1] (0.000513μM**)Value of % activation at 0.000513 uM Agonist concentration; replicate [1]Float%
33Activation at 0.000513 uM Ag [2] (0.000513μM**)Value of % activation at 0.000513 uM Agonist concentration; replicate [2]Float%
34Activation at 0.000513 uM Ag [3] (0.000513μM**)Value of % activation at 0.000513 uM Agonist concentration; replicate [3]Float%
35Activation at 0.000170 uM Ag [1] (0.00017μM**)Value of % activation at 0.000170 uM Agonist concentration; replicate [1]Float%
36Activation at 0.000170 uM Ag [2] (0.00017μM**)Value of % activation at 0.000170 uM Agonist concentration; replicate [2]Float%
37Activation at 0.000170 uM Ag [3] (0.00017μM**)Value of % activation at 0.000170 uM Agonist concentration; replicate [3]Float%
38Activation at 0.0000562 uM Ag [1] (5.62e-05μM**)Value of % activation at 0.0000562 uM Agonist concentration; replicate [1]Float%
39Activation at 0.0000562 uM Ag [2] (5.62e-05μM**)Value of % activation at 0.0000562 uM Agonist concentration; replicate [2]Float%
40Activation at 0.0000562 uM Ag [3] (5.62e-05μM**)Value of % activation at 0.0000562 uM Agonist concentration; replicate [3]Float%
41Activation at 0.0000186 uM Ag [1] (1.86e-05μM**)Value of % activation at 0.0000186 uM Agonist concentration; replicate [1]Float%
42Activation at 0.0000186 uM Ag [2] (1.86e-05μM**)Value of % activation at 0.0000186 uM Agonist concentration; replicate [2]Float%
43Activation at 0.0000186 uM Ag [3] (1.86e-05μM**)Value of % activation at 0.0000186 uM Agonist concentration; replicate [3]Float%
44Activation at 0.0000062 uM Ag [1] (6.2e-06μM**)Value of % activation at 0.0000062 uM Agonist concentration; replicate [1]Float%
45Activation at 0.0000062 uM Ag [2] (6.2e-06μM**)Value of % activation at 0.0000062 uM Agonist concentration; replicate [2]Float%
46Activation at 0.0000062 uM Ag [3] (6.2e-06μM**)Value of % activation at 0.0000062 uM Agonist concentration; replicate [3]Float%
47Activation at 0.0000020 uM Ag [1] (2e-06μM**)Value of % activation at 0.0000020 uM Agonist concentration; replicate [1]Float%
48Activation at 0.0000020 uM Ag [2] (2e-06μM**)Value of % activation at 0.0000020 uM Agonist concentration; replicate [2]Float%
49Activation at 0.0000020 uM Ag [3] (2e-06μM**)Value of % activation at 0.0000020 uM Agonist concentration; replicate [3]Float%

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: U01 AI074564

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