Name: Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based high throughput dose response assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A). ..more
Target
Tested Compounds:
Depositor Specified Assays
Show more |
| AID | Name | Type | Comment |
|---|---|---|---|
| 504326 | Luminescence-based cell-based primary high throughput screening assay to identify agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors | screening | Primary screen (OPRM1-OPRD1 agonists in singlicate) |
| 504355 | Summary of the probe development efforts to identify agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors | summary | Summary (OPRM1-OPRD1 agonists) |
| 504692 | Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based full-deck high throughput screening assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A) | screening | Counterscreen (HTR5A agonists in singlicate) |
| 504904 | Luminescence-based cell-based high throughput confirmation assay for agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors | screening | Confirmation screen (OPRM1-OPRD1 agonists in triplicate) |
| 504905 | Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based high throughput screening assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A) | screening | Counterscreen (HTR5A agonists in triplicate) |
| 651685 | On Hold | ||
| 651686 | On Hold | ||
| 651688 | On Hold | ||
| 651689 | On Hold | ||
| 651762 | On Hold | ||
| 651764 | On Hold | ||
| 651765 | On Hold | ||
| 651769 | On Hold | ||
| 651904 | On Hold |
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Lakshmi A. Devi, Mount Sinai School of Medicine
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: R03NS053751
Grant Proposal PI: Lakshmi A. Devi, Mount Sinai School of Medicine
External Assay ID: HTR5A_AG_LUMI_1536_3XEC50 DCSRUN
Name: Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based high throughput dose response assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A).
Description:
Opiates such as morphine are the choice analgesic in the treatment of chronic pain due to their potent and rapid action. However, their long-term use is limited because of the development of tolerance and dependence, as well as respiratory suppression and constipation (1). Due to their clinical importance, various strategies have been considered for making opiates more effective while curbing liabilities such as addiction. One such strategy has been to use a combination of drugs to improve the effectiveness of morphine. The OPRM1 gene encodes the mu opioid receptor, which is the primary site of action for morphine (2) and other commonly used opioid such as heroin, fentanyl, and methadone. OPRM1 activation and subsequent dissociation of the Gi/Go G-proteins results in reduction of adenylyl cyclase-mediated cAMP production (3). There are at least two other types of opioid receptors: delta (OPRD1) and kappa (OPRK1), each with a distinct pharmacologic profile. In particular, delta (OPRD1) opioid receptor ligands have been useful in enhancing morphine's potency, but the underlying molecular basis is not understood (4). It has been shown that modulation of receptor function by physical association between mu and delta opioid receptors is a potential mechanism (5). The assay provider has previously found that a combination of OPRM1 agonist with OPRD1 antagonist selectively activates the OPRM1-OPRD1 heteromer (5) and recently showed that this could be blocked by antibodies that selectively recognize the heteromer (6). Since OPRD1 antagonist have anxiogenic effects, these are not ideal as therapies. Hence, the identification of compounds that selectively activate mu-delta opioid receptor heterodimerization may have potential in the treatment of pain and alleviate unwanted effects associated with opiate use.
References:
1. Raehal KM, Bohn LM. Mu opioid receptor regulation and opiate responsiveness. AAPS J. 2005 Oct 19;7(3):E587-91.
2. Matthes H, Maldonado R, Simonin F, Valverde O, Slowe S, Kitchen I, Befort K, Dierich A, Le Meur M, Dolle P, Tzavara E, Hanoune J, Roques B, Kieffer B (1996) Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the mu-opioid-receptor gene. Nature 383:819-823.
3. Maguea SD and Blendy JAOPRM1 SNP (A118G): Involvement in disease development, treatment response, and animal models. Drug and Alcohol Dependence. 2010 May 108 (3): 172-182.
4. Traynor J, Elliot J. Delta-opioid receptor subtypes and cross talk with mu-receptors. Trends Pharmacol Sci 1993 14:84-86.
5. Gomes I, Jordan BA, Gupta A, Trapaidze N, Nagy V, Devi LA. Heterodimerization of mu and delta opioid receptors: A role in opiate synergy. J Neurosci. 2000 Nov 15;20(22):RC110.
6. Gupta, A., Mulder, J., Gomes, I., Rozenfeld, R., Bushlin, I., Ong, E., Lim, M., Maillet, E., Junek, M., Cahill, C.M., Harkany, T. Devi, L.A. Increased abundance of opioid receptor heteromers after chronic morphine administration. Science Signaling 3:ra54, 2010
Keywords:
dose response, titration, OPRM1, MOR-1, mu, delta, OPRD1, opioid, receptor, GPCR, beta-arrestin, counterscreen, secondary, triplicate, artifact, 5HTR5A, 5HTR5A, HTR5A, HTR5A, 5-HT5A, MGC138226, serotonin, serotonin receptor 5A, U2OS, PathHunter, complementation, reconstitution, enzyme, beta-gal, beta-galactosidase, lumi, luminescence, luminescent, chemiluminescence, GPCR, receptor, agonist, activator, activate, dimer, heterodimer, homodimer, pain, HTS, high throughput screen, 1536, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Lakshmi A. Devi, Mount Sinai School of Medicine
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: R03NS053751
Grant Proposal PI: Lakshmi A. Devi, Mount Sinai School of Medicine
External Assay ID: HTR5A_AG_LUMI_1536_3XEC50 DCSRUN
Name: Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based high throughput dose response assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A).
Description:
Opiates such as morphine are the choice analgesic in the treatment of chronic pain due to their potent and rapid action. However, their long-term use is limited because of the development of tolerance and dependence, as well as respiratory suppression and constipation (1). Due to their clinical importance, various strategies have been considered for making opiates more effective while curbing liabilities such as addiction. One such strategy has been to use a combination of drugs to improve the effectiveness of morphine. The OPRM1 gene encodes the mu opioid receptor, which is the primary site of action for morphine (2) and other commonly used opioid such as heroin, fentanyl, and methadone. OPRM1 activation and subsequent dissociation of the Gi/Go G-proteins results in reduction of adenylyl cyclase-mediated cAMP production (3). There are at least two other types of opioid receptors: delta (OPRD1) and kappa (OPRK1), each with a distinct pharmacologic profile. In particular, delta (OPRD1) opioid receptor ligands have been useful in enhancing morphine's potency, but the underlying molecular basis is not understood (4). It has been shown that modulation of receptor function by physical association between mu and delta opioid receptors is a potential mechanism (5). The assay provider has previously found that a combination of OPRM1 agonist with OPRD1 antagonist selectively activates the OPRM1-OPRD1 heteromer (5) and recently showed that this could be blocked by antibodies that selectively recognize the heteromer (6). Since OPRD1 antagonist have anxiogenic effects, these are not ideal as therapies. Hence, the identification of compounds that selectively activate mu-delta opioid receptor heterodimerization may have potential in the treatment of pain and alleviate unwanted effects associated with opiate use.
References:
1. Raehal KM, Bohn LM. Mu opioid receptor regulation and opiate responsiveness. AAPS J. 2005 Oct 19;7(3):E587-91.
2. Matthes H, Maldonado R, Simonin F, Valverde O, Slowe S, Kitchen I, Befort K, Dierich A, Le Meur M, Dolle P, Tzavara E, Hanoune J, Roques B, Kieffer B (1996) Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the mu-opioid-receptor gene. Nature 383:819-823.
3. Maguea SD and Blendy JAOPRM1 SNP (A118G): Involvement in disease development, treatment response, and animal models. Drug and Alcohol Dependence. 2010 May 108 (3): 172-182.
4. Traynor J, Elliot J. Delta-opioid receptor subtypes and cross talk with mu-receptors. Trends Pharmacol Sci 1993 14:84-86.
5. Gomes I, Jordan BA, Gupta A, Trapaidze N, Nagy V, Devi LA. Heterodimerization of mu and delta opioid receptors: A role in opiate synergy. J Neurosci. 2000 Nov 15;20(22):RC110.
6. Gupta, A., Mulder, J., Gomes, I., Rozenfeld, R., Bushlin, I., Ong, E., Lim, M., Maillet, E., Junek, M., Cahill, C.M., Harkany, T. Devi, L.A. Increased abundance of opioid receptor heteromers after chronic morphine administration. Science Signaling 3:ra54, 2010
Keywords:
dose response, titration, OPRM1, MOR-1, mu, delta, OPRD1, opioid, receptor, GPCR, beta-arrestin, counterscreen, secondary, triplicate, artifact, 5HTR5A, 5HTR5A, HTR5A, HTR5A, 5-HT5A, MGC138226, serotonin, serotonin receptor 5A, U2OS, PathHunter, complementation, reconstitution, enzyme, beta-gal, beta-galactosidase, lumi, luminescence, luminescent, chemiluminescence, GPCR, receptor, agonist, activator, activate, dimer, heterodimer, homodimer, pain, HTS, high throughput screen, 1536, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Protocol
Assay Overview:
The purpose of this assay is to determine whether compounds identified as active in a set of experiments entitled, "Luminescence-based cell-based primary high throughput screening assay to identify agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors" (AID 504326), that confirmed activity in a set of experiments entitled, "Luminescence-based cell-based high throughput confirmation assay for agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors" (AID 504904), and that were inactive in a set of experiments entitled, "Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based full-deck high throughput screening assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A)" (AID 504692) are nonselective GPCR activators/agonists due to activation of the HTR5A.
The assay monitors GPCR-beta-arrestin proximity using low affinity fragment complementation of beta-galactosidase (beta-gal). This assay employs U2OS cells which express HTR5A fused to a beta-gal peptide fragment (enzyme donor), and beta-arrestin fused to the complementary beta-gal fragment (enzyme acceptor). Cells are incubated with test compound, followed by measurement of well luminescence. As designed, compounds that activate HTR5A will cause beta-arrestin recruitment, resulting in reconstitution of the beta-gal holoenzyme. The reconstituted holoenzyme can then catalyze the hydrolysis of a substrate which yields a chemiluminescent signal, resulting in increased well luminescence. Serotonin (5HT) will be used as the high (100% RLU) control for agonists, and wells containing cells treated with DMSO will be used as the low (0% RLU) control. Compounds were tested in triplicate using a 10-point, 1:3 dilution series, starting at a nominal test concentration of 93 uM.
Protocol Summary:
The U2OS-HTR5A cell line was routinely cultured in T-175 sq cm flasks at 37 C and 95% relative humidity (RH). The growth media consisted of a 1:1 mixture of Ham's F-12 Nutrient Media (F-12) and Dulbecco's Modified Eagle Media (DMEM) supplemented with 10% v/v heat-inactivated certified fetal bovine serum, 25 mM HEPES, 250 ug/mL Geneticin, 250 ug/mL Hygromycin B, and 1X antibiotic mix (penicillin, streptomycin, and neomycin).
The day before the assay 1000 cells in 3 uL of cell plating media were seeded into each well of 1536 well microtiter plates and allowed to incubate at 37 C, 5% CO2, and 95 % RH for 23 hours. Next, 28 nL of test compound in DMSO, or DMSO alone were dispensed to the appropriate wells. The plates were then incubated for 3 hours at 37 C, 5% CO2, and 95 % RH. The assay was started by adding 2 uL of PathHunter TM reagent (prepared according to the manufacturer's protocol); followed by 1 hour incubation at room temperature. Well Luminescence was read on the ViewLux plate reader. The percent activation for each compound was calculated as follows:
%_Activation = ( ( RLU_Test_Compound - Median_RLU_Low_Control ) / ( Median_RLU_High_Control - Median_RLU_Low_Control ) ) * 100
Where:
High_Control is defined as wells containing cells, 5HT and DMSO.
Test_Compound is defined as wells containing cells, test compounds and DMSO.
Low_Control is defined as wells containing cells and DMSO.
For each test compound, percent activation was plotted against compound concentration. A four parameter equation describing a sigmoidal dose-response curve was then fitted with adjustable baseline using Assay Explorer software (Symyx Technologies Inc). The reported EC50 values were generated from fitted curves by solving for the X-intercept value at the 50% activation level of the Y-intercept value. In cases where the highest concentration tested (i.e. 93 uM) did not result in greater than 50% activation, the EC50 was determined manually as greater than 93 uM.
PubChem Activity Outcome and Score:
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.
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, with the most potent compounds assigned the highest activity scores.
List of Reagents:
PathHunter TM B-arrestin recruitment assay, containing the U2OS HTR5A Beta-arrestin cell line; (DiscoveRx, part 930001)
PathHunter Detection Kit (DiscoveRx, part 93-0700C3)
Ham's F-12 media (Invitrogen, part 11765-054)
DMEM media (Invitrogen, part 11995-073)
Detachin (Genlantis, part T100100)
Heat Inactivated Fetal Bovine Serum (Invitrogen, part 10082-147)
Hygromycin B (Invitrogen, part 10687-010)
Geneticin (Invitrogen, part 10131-027)
100X Penicillin-Streptomycin-Neomycin mix (Invitrogen, part 15640-055)
T-175 tissue culture flasks (Nunc, part 159910)
Agonist: Serotonin hydrochloride (5HT) (Sigma-Aldrich, part H9523)
1536-well plates (Greiner, part 789173)
The purpose of this assay is to determine whether compounds identified as active in a set of experiments entitled, "Luminescence-based cell-based primary high throughput screening assay to identify agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors" (AID 504326), that confirmed activity in a set of experiments entitled, "Luminescence-based cell-based high throughput confirmation assay for agonists of heterodimerization of the mu 1 (OPRM1) and delta 1 (OPRD1) opioid receptors" (AID 504904), and that were inactive in a set of experiments entitled, "Counterscreen for agonists of OPRM1-OPRD1 heterodimerization: luminescence-based cell-based full-deck high throughput screening assay to identify agonists of 5-hydroxytryptamine (serotonin) 5A receptor (HTR5A)" (AID 504692) are nonselective GPCR activators/agonists due to activation of the HTR5A.
The assay monitors GPCR-beta-arrestin proximity using low affinity fragment complementation of beta-galactosidase (beta-gal). This assay employs U2OS cells which express HTR5A fused to a beta-gal peptide fragment (enzyme donor), and beta-arrestin fused to the complementary beta-gal fragment (enzyme acceptor). Cells are incubated with test compound, followed by measurement of well luminescence. As designed, compounds that activate HTR5A will cause beta-arrestin recruitment, resulting in reconstitution of the beta-gal holoenzyme. The reconstituted holoenzyme can then catalyze the hydrolysis of a substrate which yields a chemiluminescent signal, resulting in increased well luminescence. Serotonin (5HT) will be used as the high (100% RLU) control for agonists, and wells containing cells treated with DMSO will be used as the low (0% RLU) control. Compounds were tested in triplicate using a 10-point, 1:3 dilution series, starting at a nominal test concentration of 93 uM.
Protocol Summary:
The U2OS-HTR5A cell line was routinely cultured in T-175 sq cm flasks at 37 C and 95% relative humidity (RH). The growth media consisted of a 1:1 mixture of Ham's F-12 Nutrient Media (F-12) and Dulbecco's Modified Eagle Media (DMEM) supplemented with 10% v/v heat-inactivated certified fetal bovine serum, 25 mM HEPES, 250 ug/mL Geneticin, 250 ug/mL Hygromycin B, and 1X antibiotic mix (penicillin, streptomycin, and neomycin).
The day before the assay 1000 cells in 3 uL of cell plating media were seeded into each well of 1536 well microtiter plates and allowed to incubate at 37 C, 5% CO2, and 95 % RH for 23 hours. Next, 28 nL of test compound in DMSO, or DMSO alone were dispensed to the appropriate wells. The plates were then incubated for 3 hours at 37 C, 5% CO2, and 95 % RH. The assay was started by adding 2 uL of PathHunter TM reagent (prepared according to the manufacturer's protocol); followed by 1 hour incubation at room temperature. Well Luminescence was read on the ViewLux plate reader. The percent activation for each compound was calculated as follows:
%_Activation = ( ( RLU_Test_Compound - Median_RLU_Low_Control ) / ( Median_RLU_High_Control - Median_RLU_Low_Control ) ) * 100
Where:
High_Control is defined as wells containing cells, 5HT and DMSO.
Test_Compound is defined as wells containing cells, test compounds and DMSO.
Low_Control is defined as wells containing cells and DMSO.
For each test compound, percent activation was plotted against compound concentration. A four parameter equation describing a sigmoidal dose-response curve was then fitted with adjustable baseline using Assay Explorer software (Symyx Technologies Inc). The reported EC50 values were generated from fitted curves by solving for the X-intercept value at the 50% activation level of the Y-intercept value. In cases where the highest concentration tested (i.e. 93 uM) did not result in greater than 50% activation, the EC50 was determined manually as greater than 93 uM.
PubChem Activity Outcome and Score:
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.
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, with the most potent compounds assigned the highest activity scores.
List of Reagents:
PathHunter TM B-arrestin recruitment assay, containing the U2OS HTR5A Beta-arrestin cell line; (DiscoveRx, part 930001)
PathHunter Detection Kit (DiscoveRx, part 93-0700C3)
Ham's F-12 media (Invitrogen, part 11765-054)
DMEM media (Invitrogen, part 11995-073)
Detachin (Genlantis, part T100100)
Heat Inactivated Fetal Bovine Serum (Invitrogen, part 10082-147)
Hygromycin B (Invitrogen, part 10687-010)
Geneticin (Invitrogen, part 10131-027)
100X Penicillin-Streptomycin-Neomycin mix (Invitrogen, part 15640-055)
T-175 tissue culture flasks (Nunc, part 159910)
Agonist: Serotonin hydrochloride (5HT) (Sigma-Aldrich, part H9523)
1536-well plates (Greiner, part 789173)
Comment
Due to the increasing size of the MLPCN compound library, this assay may have been run as two or more separate campaigns, each campaign testing a unique set of compounds. All data reported were normalized on a per-plate basis. Possible artifacts of this assay can include, but are not limited to: dust or lint located in or on wells of the microtiter plate, and compounds that modulate well luminescence. All test compound concentrations reported above and below are nominal; the specific test concentration(s) for a particular compound may vary based upon the actual sample provided by the MLSMR. The MLSMR was unable to provide all samples requested for testing.
Categorized Comment
Experiment Date: 09/22/2011
BAO: version: 1.4b1080
BAO: bioassay specification: assay stage: secondary: counter screen
BAO: bioassay specification: assay biosafety level: bsl1
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: format detail: reagent: inducer: interferon gamma
BAO: meta target: molecular target: protein target: receptor: transmembrane receptor: g protein coupled receptor
BAO: meta target: biological process target: regulation of molecular function
BAO: meta target detail: binding reporter specification: interaction: protein-protein
BAO: design: binding reporter: protein fragment complementation assay
BAO: detection technology: luminescence: chemiluminescence
Assay: Dictionary: Version: 0.1
Assay: CurveFit [1]: Equation: =( ( [Maximal Response] * [Concentration]^[Hill Slope] ) / ( [Inflection Point Concentration]^[Hill Slope] + [Concentration]^[Hill Slope] ) ) + [Baseline Response]
Assay: CurveFit [1]: Mask: Excluded Points
BAO: version: 1.4b1080
BAO: bioassay specification: assay stage: secondary: counter screen
BAO: bioassay specification: assay biosafety level: bsl1
BAO: bioassay specification: assay measurement type: endpoint assay
BAO: bioassay specification: assay readout content: assay readout method: regular screening
BAO: bioassay specification: assay readout content: content readout type: single readout
BAO: format detail: reagent: inducer: interferon gamma
BAO: meta target: molecular target: protein target: receptor: transmembrane receptor: g protein coupled receptor
BAO: meta target: biological process target: regulation of molecular function
BAO: meta target detail: binding reporter specification: interaction: protein-protein
BAO: design: binding reporter: protein fragment complementation assay
BAO: detection technology: luminescence: chemiluminescence
Assay: Dictionary: Version: 0.1
Assay: CurveFit [1]: Equation: =( ( [Maximal Response] * [Concentration]^[Hill Slope] ) / ( [Inflection Point Concentration]^[Hill Slope] + [Concentration]^[Hill Slope] ) ) + [Baseline Response]
Assay: CurveFit [1]: Mask: Excluded Points
Result Definitions
| Show more |
| TID | Name | Description | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||
| Score | The BioAssay activity ranking score |  | Integer | |||
| 1 | Qualifier | 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 concentration. | String | |||
| 2 | EC50* | The concentration at which 50 percent of the activity in the activator assay is observed; (EC50) shown in micromolar. | | Float | μM | |
| 3 | LogEC50 | Log10 of the qualified EC50 (EC50) from the activator assay in M concentration | | Float | ||
| 4 | Hill Slope | The variable HillSlope describes the steepness of the curve. This variable is called the Hill slope, the slope factor, or the Hill coefficient. If it is positive, the curve increases as X increases. If it is negative, the curve decreases as X increases. A standard sigmoid dose-response curve (previous equation) has a Hill Slope of 1.0. When HillSlope is less than 1.0, the curve is more shallow. When HillSlope is greater than 1.0, the curve is steeper. The Hill slope has no units. | | Float | ||
| 5 | Baseline Response | Adjustable baseline of the curve fit, minimal response value. | | Float | ||
| 6 | Maximal Response | The maximal or asymptotic response above the baseline as concentration increases without bound. | | Float | ||
| 7 | Inflection Point Concentration | The concentration value for the inflection point of the curve. | | Float | μM | |
| 8 | Excluded Points | Flags to indicate which of the dose-response points were excluded from analysis. (1) means the point was excluded and (0) means the point was not excluded. | String | |||
| 9 | Hill dS | The range of Y. | | Float | ||
| 10 | Chi Square | A measure for the 'goodness' of a fit. The chi-square test (Snedecor and Cochran, 1989) is used to test if a sample of data came from a population with a specific distribution. | | Float | ||
| 11 | Rsquare | This statistic measures how successful the fit explains the variation of the data; R-square is the square of the correlation between the response values and the predicted response values. | | Float | ||
| 12 | Number of DataPoints | Overall number of data points of normalized percent activation that was used for calculations (includes all concentration points); in some cases a data point can be excluded as outlier. | | Integer | ||
| 13 | Activation at 0.005 uM [1] (0.005μM**) | Value of % activation at 0.005 uM concentration; replicate [1] | | Float | % | |
| 14 | Activation at 0.005 uM [2] (0.005μM**) | Value of % activation at 0.005 uM concentration; replicate [2] | | Float | % | |
| 15 | Activation at 0.005 uM [3] (0.005μM**) | Value of % activation at 0.005 uM concentration; replicate [3] | | Float | % | |
| 16 | Activation at 0.014 uM [1] (0.014μM**) | Value of % activation at 0.014 uM concentration; replicate [1] | | Float | % | |
| 17 | Activation at 0.014 uM [2] (0.014μM**) | Value of % activation at 0.014 uM concentration; replicate [2] | | Float | % | |
| 18 | Activation at 0.014 uM [3] (0.014μM**) | Value of % activation at 0.014 uM concentration; replicate [3] | | Float | % | |
| 19 | Activation at 0.042 uM [1] (0.042μM**) | Value of % activation at 0.042 uM concentration; replicate [1] | | Float | % | |
| 20 | Activation at 0.042 uM [2] (0.042μM**) | Value of % activation at 0.042 uM concentration; replicate [2] | | Float | % | |
| 21 | Activation at 0.042 uM [3] (0.042μM**) | Value of % activation at 0.042 uM concentration; replicate [3] | | Float | % | |
| 22 | Activation at 0.127 uM [1] (0.127μM**) | Value of % activation at 0.127 uM concentration; replicate [1] | | Float | % | |
| 23 | Activation at 0.127 uM [2] (0.127μM**) | Value of % activation at 0.127 uM concentration; replicate [2] | | Float | % | |
| 24 | Activation at 0.127 uM [3] (0.127μM**) | Value of % activation at 0.127 uM concentration; replicate [3] | | Float | % | |
| 25 | Activation at 0.381 uM [1] (0.381μM**) | Value of % activation at 0.381 uM concentration; replicate [1] | | Float | % | |
| 26 | Activation at 0.381 uM [2] (0.381μM**) | Value of % activation at 0.381 uM concentration; replicate [2] | | Float | % | |
| 27 | Activation at 0.381 uM [3] (0.381μM**) | Value of % activation at 0.381 uM concentration; replicate [3] | | Float | % | |
| 28 | Activation at 1.1 uM [1] (1.1μM**) | Value of % activation at 1.1 uM concentration; replicate [1] | | Float | % | |
| 29 | Activation at 1.1 uM [2] (1.1μM**) | Value of % activation at 1.1 uM concentration; replicate [2] | | Float | % | |
| 30 | Activation at 1.1 uM [3] (1.1μM**) | Value of % activation at 1.1 uM concentration; replicate [3] | | Float | % | |
| 31 | Activation at 3.4 uM [1] (3.4μM**) | Value of % activation at 3.4 uM concentration; replicate [1] | | Float | % | |
| 32 | Activation at 3.4 uM [2] (3.4μM**) | Value of % activation at 3.4 uM concentration; replicate [2] | | Float | % | |
| 33 | Activation at 3.4 uM [3] (3.4μM**) | Value of % activation at 3.4 uM concentration; replicate [3] | | Float | % | |
| 34 | Activation at 10.3 uM [1] (10.3μM**) | Value of % activation at 10.3 uM concentration; replicate [1] | | Float | % | |
| 35 | Activation at 10.3 uM [2] (10.3μM**) | Value of % activation at 10.3 uM concentration; replicate [2] | | Float | % | |
| 36 | Activation at 10.3 uM [3] (10.3μM**) | Value of % activation at 10.3 uM concentration; replicate [3] | | Float | % | |
| 37 | Activation at 30.8 uM [1] (30.8μM**) | Value of % activation at 30.8 uM concentration; replicate [1] | | Float | % | |
| 38 | Activation at 30.8 uM [2] (30.8μM**) | Value of % activation at 30.8 uM concentration; replicate [2] | | Float | % | |
| 39 | Activation at 30.8 uM [3] (30.8μM**) | Value of % activation at 30.8 uM concentration; replicate [3] | | Float | % | |
| 40 | Activation at 92.5 uM [1] (92.5μM**) | Value of % activation at 92.5 uM concentration; replicate [1] | | Float | % | |
| 41 | Activation at 92.5 uM [2] (92.5μM**) | Value of % activation at 92.5 uM concentration; replicate [2] | | Float | % | |
| 42 | Activation at 92.5 uM [3] (92.5μM**) | Value of % activation at 92.5 uM concentration; replicate [3] | | Float | % |
* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: R03NS053751
Data Table (Concise)
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