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  • 94
    Millipore m w 146
    M W 146, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m w 146/product/Millipore
    Average 94 stars, based on 6 article reviews
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    92
    Avanti Polar s1p1 receptor antagonist w 146
    <t>S1P1</t> receptor-specific antagonist W-146 reversibly inhibits effect of RBC-conditioned (RBC-cond) perfusate on P s BSA , and S1P1-specific agonist SEW 2871 mimics effect of RBCs. A : representative data from a vessel show the stabilizing effect of RBC-conditioned
    S1p1 Receptor Antagonist W 146, supplied by Avanti Polar, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1p1 receptor antagonist w 146/product/Avanti Polar
    Average 92 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
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    w146  (Tocris)
    93
    Tocris w146
    S1PR2 stimulates PI3K to activate Akt. ( a and b ) PPC1 cells were infected with Ad-AC or Ad-GFP. After 6 h, cells were treated with the indicated dose of either ( a ) <t>W146</t> or ( b ) JTE013. After 48 h of infection, cells were analyzed by western blotting. ( c ) WT MEFs were infected with Asd-AC or Ad-GFP (MOI 100). After 6 h, cells were treated with the indicated dose of JTE013. After 48 h, cells were collected for western blotting. ( d ) PPC1 cells were transfected with nontargeting (SCR) or S1PR1–3-targeting shRNA vectors. After 6 h of transfection, cells were infected with Ad-GFP or Ad-AC. After 48 h of infection, cells were analyzed by western blotting and qRT–PCR. ( e ) DU145 EGFP and AC-EGFP cells were plated at low density in 96-well plates in the presence or absence of 1 μℳ JTE013, and analyzed by MTS assay on the indicated days. Statistical analysis was performed using a one-way ANOVA with Bonferroni correction, * P
    W146, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/w146/product/Tocris
    Average 93 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    w146 - by Bioz Stars, 2020-07
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    93
    Millipore w146
    Pharmacological activation of S1P 1 reverses the increased vascular permeability in Apom −/− mice. WT and Apom −/− mice ( n = 6) were treated with 10 mg/kg <t>W146</t> (S1P 1 antagonist) or SEW2871 (S1P 1 agonist) or saline 6.5 h before
    W146, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/w146/product/Millipore
    Average 93 stars, based on 44 article reviews
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    91
    Cayman Chemical w146
    S1P 1 antagonist <t>W146</t> increased lung RAR in ApoM −/− mice and skin RAR in WTmice. For the lung RAR, mice were treated with intranasal anti-ovalbumin IgG followed by ovalbumin with 0.5% Evans blue IV, and after 24 hrs subjected to bronchoalveolar lavage. Mice received W146 (10 μg) or vehicle intranasally concomitant with anti-ovalbumin IgG. (A) PMN counts (n=6 mice/group *p=0.04) and (B) RBC counts (n=6 mice/group *p=0.005) were performed. (C) For skin RAR, mice were injected intradermally with goat anti-ovalbumin IgG or PBS, followed by ovalbumin and Evan’s blue IV, and mice killed after 4hrs. Some mice received intradermal W146 (10 μg) with anti-ovalbumin IgG. A representative skin sample after RAR with intradermal W146 or vehicle is shown. (D) Skin RAR was assessed by measuring the diameter of the major and minor axis and determining area of EB leakage at the injection site (n=6; *p=0.006), and (E) the weight of 6 mm punch biopsies (n=11–12;*p=0.02).
    W146, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 91/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/w146/product/Cayman Chemical
    Average 91 stars, based on 94 article reviews
    Price from $9.99 to $1999.99
    w146 - by Bioz Stars, 2020-07
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    91
    R&D Systems w146
    S1P 1 antagonist <t>W146</t> increased lung RAR in ApoM −/− mice and skin RAR in WTmice. For the lung RAR, mice were treated with intranasal anti-ovalbumin IgG followed by ovalbumin with 0.5% Evans blue IV, and after 24 hrs subjected to bronchoalveolar lavage. Mice received W146 (10 μg) or vehicle intranasally concomitant with anti-ovalbumin IgG. (A) PMN counts (n=6 mice/group *p=0.04) and (B) RBC counts (n=6 mice/group *p=0.005) were performed. (C) For skin RAR, mice were injected intradermally with goat anti-ovalbumin IgG or PBS, followed by ovalbumin and Evan’s blue IV, and mice killed after 4hrs. Some mice received intradermal W146 (10 μg) with anti-ovalbumin IgG. A representative skin sample after RAR with intradermal W146 or vehicle is shown. (D) Skin RAR was assessed by measuring the diameter of the major and minor axis and determining area of EB leakage at the injection site (n=6; *p=0.006), and (E) the weight of 6 mm punch biopsies (n=11–12;*p=0.02).
    W146, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/w146/product/R&D Systems
    Average 91 stars, based on 2 article reviews
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    w146 - by Bioz Stars, 2020-07
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    91
    Santa Cruz Biotechnology w146
    S1P 1 antagonist <t>W146</t> increased lung RAR in ApoM −/− mice and skin RAR in WTmice. For the lung RAR, mice were treated with intranasal anti-ovalbumin IgG followed by ovalbumin with 0.5% Evans blue IV, and after 24 hrs subjected to bronchoalveolar lavage. Mice received W146 (10 μg) or vehicle intranasally concomitant with anti-ovalbumin IgG. (A) PMN counts (n=6 mice/group *p=0.04) and (B) RBC counts (n=6 mice/group *p=0.005) were performed. (C) For skin RAR, mice were injected intradermally with goat anti-ovalbumin IgG or PBS, followed by ovalbumin and Evan’s blue IV, and mice killed after 4hrs. Some mice received intradermal W146 (10 μg) with anti-ovalbumin IgG. A representative skin sample after RAR with intradermal W146 or vehicle is shown. (D) Skin RAR was assessed by measuring the diameter of the major and minor axis and determining area of EB leakage at the injection site (n=6; *p=0.006), and (E) the weight of 6 mm punch biopsies (n=11–12;*p=0.02).
    W146, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/w146/product/Santa Cruz Biotechnology
    Average 91 stars, based on 1 article reviews
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    86
    Cayman Chemical s1p receptors antagonists w146
    Receptors involved in the cooperative effect. AVICs were pre-treated with the indicated drugs, activated for 12 h, and cell lysates and supernatants were analyzed by Western blot and ELISA, respectively. A) Representative immunoblots with densitometry data demonstrate inhibition of the cooperative effect on COX-2 and ICAM-1 expression (100% value) using <t>S1P</t> 1/3 antagonists (n = 6–12). B) Silencing S1P 1/3 attenuated the cooperative effect (n = 3 control AVIC). Scramble, siRNA control; vehicle, 0.1% DEPC. C) Immunoblots showed cooperation between FTY720 and LPS (n = 3). D) ELISA quantification of sICAM-1 levels show inhibition by S1P 1/3 antagonists (n = 6–10). E) Immunoblots demonstrate inhibition of the cooperative effect by TLR4 antagonists (n = 3). Cay indicates 5 µM CAY10614; CLI, 3 µM CLI-095; FTY, 1 µM FTY720; JTE, 10 µM JTE-013; S+L, S1P+LPS; PTX, 100 ng/ml pertussis toxin; R, resting; Sur, 10 µM suramin; <t>W146,</t> 10 µM W146. * p
    S1p Receptors Antagonists W146, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1p receptors antagonists w146/product/Cayman Chemical
    Average 86 stars, based on 1 article reviews
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    93
    Cayman Chemical s1pr1 antagonist w146
    Receptors involved in the cooperative effect. AVICs were pre-treated with the indicated drugs, activated for 12 h, and cell lysates and supernatants were analyzed by Western blot and ELISA, respectively. A) Representative immunoblots with densitometry data demonstrate inhibition of the cooperative effect on COX-2 and ICAM-1 expression (100% value) using <t>S1P</t> 1/3 antagonists (n = 6–12). B) Silencing S1P 1/3 attenuated the cooperative effect (n = 3 control AVIC). Scramble, siRNA control; vehicle, 0.1% DEPC. C) Immunoblots showed cooperation between FTY720 and LPS (n = 3). D) ELISA quantification of sICAM-1 levels show inhibition by S1P 1/3 antagonists (n = 6–10). E) Immunoblots demonstrate inhibition of the cooperative effect by TLR4 antagonists (n = 3). Cay indicates 5 µM CAY10614; CLI, 3 µM CLI-095; FTY, 1 µM FTY720; JTE, 10 µM JTE-013; S+L, S1P+LPS; PTX, 100 ng/ml pertussis toxin; R, resting; Sur, 10 µM suramin; <t>W146,</t> 10 µM W146. * p
    S1pr1 Antagonist W146, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1pr1 antagonist w146/product/Cayman Chemical
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    s1pr1 antagonist w146 - by Bioz Stars, 2020-07
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    84
    Avanti Polar s1pr1 w146
    Receptors involved in the cooperative effect. AVICs were pre-treated with the indicated drugs, activated for 12 h, and cell lysates and supernatants were analyzed by Western blot and ELISA, respectively. A) Representative immunoblots with densitometry data demonstrate inhibition of the cooperative effect on COX-2 and ICAM-1 expression (100% value) using <t>S1P</t> 1/3 antagonists (n = 6–12). B) Silencing S1P 1/3 attenuated the cooperative effect (n = 3 control AVIC). Scramble, siRNA control; vehicle, 0.1% DEPC. C) Immunoblots showed cooperation between FTY720 and LPS (n = 3). D) ELISA quantification of sICAM-1 levels show inhibition by S1P 1/3 antagonists (n = 6–10). E) Immunoblots demonstrate inhibition of the cooperative effect by TLR4 antagonists (n = 3). Cay indicates 5 µM CAY10614; CLI, 3 µM CLI-095; FTY, 1 µM FTY720; JTE, 10 µM JTE-013; S+L, S1P+LPS; PTX, 100 ng/ml pertussis toxin; R, resting; Sur, 10 µM suramin; <t>W146,</t> 10 µM W146. * p
    S1pr1 W146, supplied by Avanti Polar, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1pr1 w146/product/Avanti Polar
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    85
    Cayman Chemical s1p1 antagonist w146
    Effect of S1P agonist and/or <t>S1P1</t> antagonist on urinary volume (U·V) and sodium excretion (U Na ·V). A : U·V. B : U Na ·V. FTY720, S1P agonist; <t>W146,</t> S1P1 antagonist; C, control with vehicle infusion; T, treatment with reagent
    S1p1 Antagonist W146, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1p1 antagonist w146/product/Cayman Chemical
    Average 85 stars, based on 2 article reviews
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    Image Search Results


    S1P1 receptor-specific antagonist W-146 reversibly inhibits effect of RBC-conditioned (RBC-cond) perfusate on P s BSA , and S1P1-specific agonist SEW 2871 mimics effect of RBCs. A : representative data from a vessel show the stabilizing effect of RBC-conditioned

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Erythrocyte-derived sphingosine-1-phosphate stabilizes basal hydraulic conductivity and solute permeability in rat microvessels

    doi: 10.1152/ajpheart.00181.2012

    Figure Lengend Snippet: S1P1 receptor-specific antagonist W-146 reversibly inhibits effect of RBC-conditioned (RBC-cond) perfusate on P s BSA , and S1P1-specific agonist SEW 2871 mimics effect of RBCs. A : representative data from a vessel show the stabilizing effect of RBC-conditioned

    Article Snippet: Stock solution of S1P1 receptor antagonist W-146 (Avanti Polar Lipids,) was prepared at 5 mM with 2% 2-hydroxypropyl-β-cyclodextrin in Ringer; W-146 was diluted to working concentration (10 μM and 0.004% 2-hydroxypropyl-β-cyclodextrin in BSA/Ringer) with or without added fluorescent solute immediately before each experiment.

    Techniques:

    Effects of S1P and S1P 1 receptor antagonist W146 on sGAGs (HS and CS). Coverage analysis of HS ( A and B ) and CS ( C and D ) on rat fat-pad endothelial cells (RFPECs), which were treated with p-DMEM or f-DMEM in the presence of S1P ( A and C ), or treated

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Sphingosine-1-phosphate protects endothelial glycocalyx by inhibiting syndecan-1 shedding

    doi: 10.1152/ajpheart.00687.2013

    Figure Lengend Snippet: Effects of S1P and S1P 1 receptor antagonist W146 on sGAGs (HS and CS). Coverage analysis of HS ( A and B ) and CS ( C and D ) on rat fat-pad endothelial cells (RFPECs), which were treated with p-DMEM or f-DMEM in the presence of S1P ( A and C ), or treated

    Article Snippet: The effects of S1P and W146 on both HS and CS were further examined as shown in .

    Techniques:

    Defects in stabilization of VE-cadherin-mediated cellular junctions in Memo knock-down endothelial cells. A , Effects of S1P or S1P receptor blocker on junctional localization of VE-cadherin in control and Memo KD HUVECs. Monolayers of HUVECs were cultured for 6 h in full media or serum-free media with or without VPC23019 (1 μM) or S1P (1 μM), then fixed and stained for VE-cadherin. Scale bar, 40 μm. B–C , Sprout formation from multicellular spheroids generated from control and Memo KD HUVECs. Sprouting was compared in the absence and presence of the S1PR1-specific agonist, SEW2871 (100 nM) or the S1PR1-specific antagonist, W146 (10 μM). Representative images (B) and quantified results (C) are shown. The numbers of multicellular spheroids used for each condition are indicated at the bottom of (C). Scale bar in (B), 100 μm. Data in (C) are presented as means ± S.D. of the scores for each multicellular spheroid. *, p

    Journal: PLoS ONE

    Article Title: Memo Has a Novel Role in S1P Signaling and Crucial for Vascular Development

    doi: 10.1371/journal.pone.0094114

    Figure Lengend Snippet: Defects in stabilization of VE-cadherin-mediated cellular junctions in Memo knock-down endothelial cells. A , Effects of S1P or S1P receptor blocker on junctional localization of VE-cadherin in control and Memo KD HUVECs. Monolayers of HUVECs were cultured for 6 h in full media or serum-free media with or without VPC23019 (1 μM) or S1P (1 μM), then fixed and stained for VE-cadherin. Scale bar, 40 μm. B–C , Sprout formation from multicellular spheroids generated from control and Memo KD HUVECs. Sprouting was compared in the absence and presence of the S1PR1-specific agonist, SEW2871 (100 nM) or the S1PR1-specific antagonist, W146 (10 μM). Representative images (B) and quantified results (C) are shown. The numbers of multicellular spheroids used for each condition are indicated at the bottom of (C). Scale bar in (B), 100 μm. Data in (C) are presented as means ± S.D. of the scores for each multicellular spheroid. *, p

    Article Snippet: VPC23019 and (R)-W146 were obtained from Avanti Polar Lipids (Alabaster, AL).

    Techniques: Cell Culture, Staining, Generated

    DH-S1P enhances endothelial barrier in an S1P1 dependent manner . Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A , EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B , EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C , EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D , EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B , monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D , ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.

    Journal: Lipids in Health and Disease

    Article Title: S1P, dihydro-S1P and C24:1-ceramide levels in the HDL-containing fraction of serum inversely correlate with occurrence of ischemic heart disease

    doi: 10.1186/1476-511X-10-70

    Figure Lengend Snippet: DH-S1P enhances endothelial barrier in an S1P1 dependent manner . Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A , EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B , EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C , EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D , EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B , monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D , ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.

    Article Snippet: For experiments evaluating the effects of the S1P1 antagonist W146 (Avanti Polar Lipids) and the S1P1/S1P3 antagonist VPC23019 (Avanti Polar Lipids) on TEER, antagonist stocks (1 mM in 5% acidified DMSO, 4 mg/ml BSA) were diluted 1:100 into the conditioned EBM at the same time that S1P or HDL was added.

    Techniques: Incubation, Concentration Assay

    DH-S1P enhances endothelial cell motility in an S1P1 dependent manner . EC monolayers were wounded with a burst of high electrical current as described previously [ 5 ] and the culture medium then supplemented with DH-S1P (1 μM) ( A ), or S1P (1 μM) ( B ) in the presence or absence of the S1P1 antagonist W146 (10 μM in DMSO vehicle). The migration of cells into the wounded areas was measured in real-time by electrical impedance. As a control in both experiments the medium was supplemented with delipidated BSA (Control) in PBS. Electrical impedance data are normalized to baseline following wounding. The data depicted are representative of two independent experiments and traces represent averages of two replicates per condition.

    Journal: Lipids in Health and Disease

    Article Title: S1P, dihydro-S1P and C24:1-ceramide levels in the HDL-containing fraction of serum inversely correlate with occurrence of ischemic heart disease

    doi: 10.1186/1476-511X-10-70

    Figure Lengend Snippet: DH-S1P enhances endothelial cell motility in an S1P1 dependent manner . EC monolayers were wounded with a burst of high electrical current as described previously [ 5 ] and the culture medium then supplemented with DH-S1P (1 μM) ( A ), or S1P (1 μM) ( B ) in the presence or absence of the S1P1 antagonist W146 (10 μM in DMSO vehicle). The migration of cells into the wounded areas was measured in real-time by electrical impedance. As a control in both experiments the medium was supplemented with delipidated BSA (Control) in PBS. Electrical impedance data are normalized to baseline following wounding. The data depicted are representative of two independent experiments and traces represent averages of two replicates per condition.

    Article Snippet: For experiments evaluating the effects of the S1P1 antagonist W146 (Avanti Polar Lipids) and the S1P1/S1P3 antagonist VPC23019 (Avanti Polar Lipids) on TEER, antagonist stocks (1 mM in 5% acidified DMSO, 4 mg/ml BSA) were diluted 1:100 into the conditioned EBM at the same time that S1P or HDL was added.

    Techniques: Migration

    S1PR2 stimulates PI3K to activate Akt. ( a and b ) PPC1 cells were infected with Ad-AC or Ad-GFP. After 6 h, cells were treated with the indicated dose of either ( a ) W146 or ( b ) JTE013. After 48 h of infection, cells were analyzed by western blotting. ( c ) WT MEFs were infected with Asd-AC or Ad-GFP (MOI 100). After 6 h, cells were treated with the indicated dose of JTE013. After 48 h, cells were collected for western blotting. ( d ) PPC1 cells were transfected with nontargeting (SCR) or S1PR1–3-targeting shRNA vectors. After 6 h of transfection, cells were infected with Ad-GFP or Ad-AC. After 48 h of infection, cells were analyzed by western blotting and qRT–PCR. ( e ) DU145 EGFP and AC-EGFP cells were plated at low density in 96-well plates in the presence or absence of 1 μℳ JTE013, and analyzed by MTS assay on the indicated days. Statistical analysis was performed using a one-way ANOVA with Bonferroni correction, * P

    Journal: Oncogenesis

    Article Title: Acid ceramidase induces sphingosine kinase 1/S1P receptor 2-mediated activation of oncogenic Akt signaling

    doi: 10.1038/oncsis.2013.14

    Figure Lengend Snippet: S1PR2 stimulates PI3K to activate Akt. ( a and b ) PPC1 cells were infected with Ad-AC or Ad-GFP. After 6 h, cells were treated with the indicated dose of either ( a ) W146 or ( b ) JTE013. After 48 h of infection, cells were analyzed by western blotting. ( c ) WT MEFs were infected with Asd-AC or Ad-GFP (MOI 100). After 6 h, cells were treated with the indicated dose of JTE013. After 48 h, cells were collected for western blotting. ( d ) PPC1 cells were transfected with nontargeting (SCR) or S1PR1–3-targeting shRNA vectors. After 6 h of transfection, cells were infected with Ad-GFP or Ad-AC. After 48 h of infection, cells were analyzed by western blotting and qRT–PCR. ( e ) DU145 EGFP and AC-EGFP cells were plated at low density in 96-well plates in the presence or absence of 1 μℳ JTE013, and analyzed by MTS assay on the indicated days. Statistical analysis was performed using a one-way ANOVA with Bonferroni correction, * P

    Article Snippet: Reagents used include: SKI–II, Docetaxel (Thermo Fisher, Lafayette, CO, USA), LY294002, Wortmannin, AktX (Cayman Chemical, Ann Arbor, MI, USA), W146, JTE013, NF023 (Tocris, Bristol, UK), Perifosine (BioVision, Milpitas, CA, USA) and pertussis toxin (Sigma Aldrich, St Louis, MO, USA).

    Techniques: Infection, Western Blot, Transfection, shRNA, Quantitative RT-PCR, MTS Assay

    The migration of integrin alpha-6-positive germ cell precursors towards sphingosine-1-phosphate is mediated by the G-protein–coupled receptor S1PR1 and integrin alpha-dependent binding to laminin. ( a ) Migration assay of ALDH-positive cells in response to different concentrations of S1P, as indicated. No stimulant was added to control wells. Sorted cells were added to the upper chamber of a Transwell system coated with laminin, and after 2 h, migrated cells in the lower chamber were counted. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls ( n =6). Statistical analysis was performed using Student's t -test. ** P ≤0.05 (medium significance compared with control). ( b ) Migration assay of ALDH-positive/Integrin-alpha-6-positive cells in the presence of 2 μm S1P with or without inhibitors of S1PR signalling. S1PR1 antagonist W146 (10 μM), PI3K-inhibitor Wortmannin (0.1 μM) or Rac1-inhibitor NSC (100 μM) were added as indicated. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls. ( c ) Migration assay of ALDH-positive/integrin alpha-6-positive cells on uncoated plastic or laminin-coated Transwells with or without 2 μm S1P. Anti-integrin alpha-6 (5 μg ml −1 ) or isotype control antibody was added as indicated. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls on laminin. Error bars represent the standard deviation for each average ( n =6) Statistical analysis was performed using Student's t -test. ** P ≤0.05 (medium significance).

    Journal: Nature Communications

    Article Title: Migration of germline progenitor cells is directed by sphingosine-1-phosphate signalling in a basal chordate

    doi: 10.1038/ncomms9565

    Figure Lengend Snippet: The migration of integrin alpha-6-positive germ cell precursors towards sphingosine-1-phosphate is mediated by the G-protein–coupled receptor S1PR1 and integrin alpha-dependent binding to laminin. ( a ) Migration assay of ALDH-positive cells in response to different concentrations of S1P, as indicated. No stimulant was added to control wells. Sorted cells were added to the upper chamber of a Transwell system coated with laminin, and after 2 h, migrated cells in the lower chamber were counted. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls ( n =6). Statistical analysis was performed using Student's t -test. ** P ≤0.05 (medium significance compared with control). ( b ) Migration assay of ALDH-positive/Integrin-alpha-6-positive cells in the presence of 2 μm S1P with or without inhibitors of S1PR signalling. S1PR1 antagonist W146 (10 μM), PI3K-inhibitor Wortmannin (0.1 μM) or Rac1-inhibitor NSC (100 μM) were added as indicated. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls. ( c ) Migration assay of ALDH-positive/integrin alpha-6-positive cells on uncoated plastic or laminin-coated Transwells with or without 2 μm S1P. Anti-integrin alpha-6 (5 μg ml −1 ) or isotype control antibody was added as indicated. Data are expressed as fold changes of numbers of migrated cells, normalized to unstimulated controls on laminin. Error bars represent the standard deviation for each average ( n =6) Statistical analysis was performed using Student's t -test. ** P ≤0.05 (medium significance).

    Article Snippet: Small-molecule inhibitor treatment Animals were incubated in 30 ml of sea water containing 1 μM Sphingosine Kinase Inhibitor (A) SKI-I (Abcam), 14 μM Sphingosine Kinase Inhibitor (B) CAS 1177741-83-1 (Millipore 567731) or 10 μM S1PR1 antagonist W146 (Tocris).

    Techniques: Migration, Binding Assay, Standard Deviation

    Pharmacological activation of S1P 1 reverses the increased vascular permeability in Apom −/− mice. WT and Apom −/− mice ( n = 6) were treated with 10 mg/kg W146 (S1P 1 antagonist) or SEW2871 (S1P 1 agonist) or saline 6.5 h before

    Journal: The FASEB Journal

    Article Title: Impaired endothelial barrier function in apolipoprotein M–deficient mice is dependent on sphingosine-1-phosphate receptor 1

    doi: 10.1096/fj.201500064

    Figure Lengend Snippet: Pharmacological activation of S1P 1 reverses the increased vascular permeability in Apom −/− mice. WT and Apom −/− mice ( n = 6) were treated with 10 mg/kg W146 (S1P 1 antagonist) or SEW2871 (S1P 1 agonist) or saline 6.5 h before

    Article Snippet: Female mice, 17–19 wk old, were injected once with SEW2871 (10 mg/kg i.p., S3944; Sigma-Aldrich) or saline 6.5 h before assessment of vascular permeability or twice with W146 (10 mg/kg i.p., W1020; Sigma-Aldrich) 6.5 and 2.5 h before assessment of vascular permeability, respectively.

    Techniques: Activation Assay, Permeability, Mouse Assay

    S1P 1 antagonist W146 increased lung RAR in ApoM −/− mice and skin RAR in WTmice. For the lung RAR, mice were treated with intranasal anti-ovalbumin IgG followed by ovalbumin with 0.5% Evans blue IV, and after 24 hrs subjected to bronchoalveolar lavage. Mice received W146 (10 μg) or vehicle intranasally concomitant with anti-ovalbumin IgG. (A) PMN counts (n=6 mice/group *p=0.04) and (B) RBC counts (n=6 mice/group *p=0.005) were performed. (C) For skin RAR, mice were injected intradermally with goat anti-ovalbumin IgG or PBS, followed by ovalbumin and Evan’s blue IV, and mice killed after 4hrs. Some mice received intradermal W146 (10 μg) with anti-ovalbumin IgG. A representative skin sample after RAR with intradermal W146 or vehicle is shown. (D) Skin RAR was assessed by measuring the diameter of the major and minor axis and determining area of EB leakage at the injection site (n=6; *p=0.006), and (E) the weight of 6 mm punch biopsies (n=11–12;*p=0.02).

    Journal: Arthritis & rheumatology (Hoboken, N.J.)

    Article Title: Sphingosine -1 Phosphate Receptor-1 signaling maintains endothelial cell barrier function and protects against immune complex-induced vascular injury

    doi: 10.1002/art.40558

    Figure Lengend Snippet: S1P 1 antagonist W146 increased lung RAR in ApoM −/− mice and skin RAR in WTmice. For the lung RAR, mice were treated with intranasal anti-ovalbumin IgG followed by ovalbumin with 0.5% Evans blue IV, and after 24 hrs subjected to bronchoalveolar lavage. Mice received W146 (10 μg) or vehicle intranasally concomitant with anti-ovalbumin IgG. (A) PMN counts (n=6 mice/group *p=0.04) and (B) RBC counts (n=6 mice/group *p=0.005) were performed. (C) For skin RAR, mice were injected intradermally with goat anti-ovalbumin IgG or PBS, followed by ovalbumin and Evan’s blue IV, and mice killed after 4hrs. Some mice received intradermal W146 (10 μg) with anti-ovalbumin IgG. A representative skin sample after RAR with intradermal W146 or vehicle is shown. (D) Skin RAR was assessed by measuring the diameter of the major and minor axis and determining area of EB leakage at the injection site (n=6; *p=0.006), and (E) the weight of 6 mm punch biopsies (n=11–12;*p=0.02).

    Article Snippet: Similar to lung RAR, there was no evidence that ApoM deficiency increased skin RAR, and intradermal injection of low doses of W146 alone did not increase EB leak ( ).

    Techniques: Mouse Assay, Injection

    Receptors involved in the cooperative effect. AVICs were pre-treated with the indicated drugs, activated for 12 h, and cell lysates and supernatants were analyzed by Western blot and ELISA, respectively. A) Representative immunoblots with densitometry data demonstrate inhibition of the cooperative effect on COX-2 and ICAM-1 expression (100% value) using S1P 1/3 antagonists (n = 6–12). B) Silencing S1P 1/3 attenuated the cooperative effect (n = 3 control AVIC). Scramble, siRNA control; vehicle, 0.1% DEPC. C) Immunoblots showed cooperation between FTY720 and LPS (n = 3). D) ELISA quantification of sICAM-1 levels show inhibition by S1P 1/3 antagonists (n = 6–10). E) Immunoblots demonstrate inhibition of the cooperative effect by TLR4 antagonists (n = 3). Cay indicates 5 µM CAY10614; CLI, 3 µM CLI-095; FTY, 1 µM FTY720; JTE, 10 µM JTE-013; S+L, S1P+LPS; PTX, 100 ng/ml pertussis toxin; R, resting; Sur, 10 µM suramin; W146, 10 µM W146. * p

    Journal: PLoS ONE

    Article Title: Synergy between Sphingosine 1-Phosphate and Lipopolysaccharide Signaling Promotes an Inflammatory, Angiogenic and Osteogenic Response in Human Aortic Valve Interstitial Cells

    doi: 10.1371/journal.pone.0109081

    Figure Lengend Snippet: Receptors involved in the cooperative effect. AVICs were pre-treated with the indicated drugs, activated for 12 h, and cell lysates and supernatants were analyzed by Western blot and ELISA, respectively. A) Representative immunoblots with densitometry data demonstrate inhibition of the cooperative effect on COX-2 and ICAM-1 expression (100% value) using S1P 1/3 antagonists (n = 6–12). B) Silencing S1P 1/3 attenuated the cooperative effect (n = 3 control AVIC). Scramble, siRNA control; vehicle, 0.1% DEPC. C) Immunoblots showed cooperation between FTY720 and LPS (n = 3). D) ELISA quantification of sICAM-1 levels show inhibition by S1P 1/3 antagonists (n = 6–10). E) Immunoblots demonstrate inhibition of the cooperative effect by TLR4 antagonists (n = 3). Cay indicates 5 µM CAY10614; CLI, 3 µM CLI-095; FTY, 1 µM FTY720; JTE, 10 µM JTE-013; S+L, S1P+LPS; PTX, 100 ng/ml pertussis toxin; R, resting; Sur, 10 µM suramin; W146, 10 µM W146. * p

    Article Snippet: In pharmacological studies, cells were pre-treated for 30 min with either S1P receptors antagonists W146 (Cayman Chem., Ann Arbor, MI); VPC 23019 (Avanti Polar Lipids, Alabaster, AL); JTE-013 (Tocris, Bristol, UK); suramin (Biomol, Santa Fe, NM), or TLR4 signaling antagonists CLI-095 (InvivoGen, San Diego, CA); CAY10614 (Cayman Chem., Ann Arbor, MI) or signaling cascades inhibitors NF-κB SN50, ALLN, SB203580, and GF109203X (Calbiochem, Darmstadt, Germany); PD98059 (Tocris, Bristol, UK); pertussis toxin (PTX) and SP600125 (Sigma, St. Louis, MO).

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Inhibition, Expressing

    Effect of S1P agonist and/or S1P1 antagonist on urinary volume (U·V) and sodium excretion (U Na ·V). A : U·V. B : U Na ·V. FTY720, S1P agonist; W146, S1P1 antagonist; C, control with vehicle infusion; T, treatment with reagent

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: A novel lipid natriuretic factor in the renal medulla: sphingosine-1-phosphate

    doi: 10.1152/ajprenal.00014.2011

    Figure Lengend Snippet: Effect of S1P agonist and/or S1P1 antagonist on urinary volume (U·V) and sodium excretion (U Na ·V). A : U·V. B : U Na ·V. FTY720, S1P agonist; W146, S1P1 antagonist; C, control with vehicle infusion; T, treatment with reagent

    Article Snippet: Group 2 : infusion of S1P1 antagonist W146 (Cayman Chemical) at 4.5, 9, and 18 μg·kg−1 ·h−1 .

    Techniques: