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  • 93
    Millipore jte 013
    <t>JTE-013</t> decreased the incidence of diabetes. JTE-013 (4 mg/kg) or saline was intraperitoneally injected for 6 days (one shot prior to STZ and five shots with a high dose of STZ (50 mg/kg)). (A) Changes in blood glucose levels of randomly fed mice with/without
    Jte 013, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cayman Chemical jte 013
    Effect of S1PR2 activation on CCA cell migration. Rat BDEsp-TDE H10 cells, human HuCCT1 cells, and CCLP1 cells were plated on six-well plates until confluent. Cells were scratched to simulate a wound and images were recorded as 0 hours. Cells were pretreated with <t>JTE-013</t> (10 μM) for 1 hour, then treated with TCA (100 μM) or S1P (100 nM) for 48 hours. Images of wound areas were recorded as described in Materials and Methods. The area of wound was quantified using IPLab4.0. Relative wound closure was calculated. (A) Rat BDEsp-TDE H10 cells and human HuCCT1 cells. (B) Human CCLP1 cells. * P
    Jte 013, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 92/100, based on 196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Tocris jte 013
    S1PR2 mediates Nogo-A-Δ20- and myelin-induced inhibition of cell spreading and neurite outgrowth. (A,C) Representative pictures of 3T3 fibroblasts treated with <t>JTE-013</t> or vehicle (DMSO) (A), or stably carrying a S1pr2 shRNA (sh- S1pr2 ) or empty vector (sh-Vec) construct (C) and plated on control, Nogo-A-Δ20 or myelin substrates. (B,D) Cell spreading quantification of (A) and (C). (E) Representative pictures of MEFs isolated from WT or S1PR2 −/− mice and plated on control, Nogo-A- Δ20, or myelin substrates. (F) Cell spreading quantification of (E). Cells were stained with Alexa488-conjugated Phalloidin in (A, C, and E). (G,I) Representative pictures of P5–8 cerebellar granule neurons treated with JTE-013 or DMSO (G), or isolated from S1PR2 −/− or WT mice (I) and plated on PLL (ctrl), Nogo-A-Δ20 or myelin substrates. (H,J) Normalized mean neurite length per cell quantification of (G) and (I). Neurons were stained with βIII-Tubulin in (G) and (I). Data shown are means ± SEM ( n = 3–6 experiments; * p
    Jte 013, supplied by Tocris, used in various techniques. Bioz Stars score: 98/100, based on 190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Selleck Chemicals s1p2 antagonist jte013
    The relationship between Foxo3a and <t>S1PR2</t> in <t>PBMCs</t> (A) and (B) PBMCs were pre-treated with LY294002 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α I and IFN γ were detected by RT-PCR. (C) and (D) PBMCs were pre-treated with SC79 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. (E) and (F) PBMCs were treated with LPS and JET-013 for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. *, p
    S1p2 Antagonist Jte013, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Cayman Chemical s1pr2 antagonist jte 013
    Rho GTPase activation. A , hMSC-TERT cells were treated with either BASE with vehicle or S1P agonist or osteoclast conditioned medium ( OC CM ) combined with either vehicle or a S1P receptor antagonists (S1PR1 and <t>S1PR2</t> antagonists together) (W123 and <t>JTE-013)</t>
    S1pr2 Antagonist Jte 013, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cambridge Bioscience jte 013
    Effect of CM from MDA-MB-231 cells on ERK-1/2 activation and DNA synthesis in MEFs MEFs quiescent for 24 hr were treated with non-conditioned medium (NCM) or conditioned medium (CM) from MDA-MB-231 cells for 10 min. MEFs were pre-treated with and without <t>JTE-013</t> (10 μM) for 15 min prior to addition of NCM or CM. MEFs were also treated with CYM5520 (10 μM) in the DNA synthesis experiments. CM was also isolated from MDA-MB-231 cells treated with scrambled or S1P 2 siRNA for 24 h. (A) Western blot showing the effect of CM from MDA-MB-231 cells and JTE-013 on ERK-1/2 activation in MEFs. (B) Bar graph showing the effect of NCM, CM from MDA-MB-231 cells or CYM5520 on [ 3 H]-thymidine incorporation into DNA synthesis in MEFs. Results are expressed as means +/- SEM for n=3. (C) Western blot showing the effect of CM on ERK-1/2 activation in MEFs. CM was isolated from MDA-MB-231 cells that had been treated with either scrambled siRNA (200 nM) or S1P 2 siRNA (100 or 200 nM). Also shown is the effect of S1P 2 siRNA on S1P 2 expression levels in MDA-MB-231 cells. (D) Western blot showing that neither P-ERK-1/2 nor ERK-2 is released from MDA-MB-231 cells into CM as evidenced by their absence in the PPT. (E) Western blot showing the lack of effect of CM from MDA-MB-453 cells on ERK-1/2 activation in MEFs. Blots were probed with anti-phospho ERK-1/2 and anti-S1P 2 antibodies. (A, C, D, E). Blots were re-probed with anti-actin antibody to ensure equal protein loading (A, C, E). Results (A, C, D, E) are representative of 3 independent experiments.
    Jte 013, supplied by Cambridge Bioscience, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology jte 013
    Effect of S1PR1-3 inhibition on angiogenesis in vitro (A) Representative images of the migration, invasion and tube formation assays. Endothelial cells were stimulated with CM collected from the ovarian cancer cells precultured with VPC23019 (300nM) or <t>JTE-013</t> (1μM). Migrated cells, invaded cells and tube like structures were photographed. (B) Statistical analysis of the migrated cells, invaded cells and tube like structures. (C) Effect of VPC23019 and JTE-013 on the VEGF, IL-8 and IL-6 secretion of ovarian cancer cells. All experiments were repeated three times, with three replicates in each group ( * p
    Jte 013, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cayman Chemical s1p receptor inhibitors jte013
    Effect of S1PR1-3 inhibition on angiogenesis in vitro (A) Representative images of the migration, invasion and tube formation assays. Endothelial cells were stimulated with CM collected from the ovarian cancer cells precultured with VPC23019 (300nM) or <t>JTE-013</t> (1μM). Migrated cells, invaded cells and tube like structures were photographed. (B) Statistical analysis of the migrated cells, invaded cells and tube like structures. (C) Effect of VPC23019 and JTE-013 on the VEGF, IL-8 and IL-6 secretion of ovarian cancer cells. All experiments were repeated three times, with three replicates in each group ( * p
    S1p Receptor Inhibitors Jte013, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Biomol GmbH s1pr2 4 antagonist jte 013
    S1PR4 on DC conveys ACM-dependent suppression of cytotoxicity. (A–D) T cells were co-cultured with control or ACM-primed autologous DC with or without the <t>S1PR2/4</t> antagonist <t>JTE-013</t> or the S1PR1/3 antagonist VPC23019. (A) Cytotoxicity induced
    S1pr2 4 Antagonist Jte 013, supplied by Biomol GmbH, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Tocris inhibitors jte013
    S1PR4 on DC conveys ACM-dependent suppression of cytotoxicity. (A–D) T cells were co-cultured with control or ACM-primed autologous DC with or without the <t>S1PR2/4</t> antagonist <t>JTE-013</t> or the S1PR1/3 antagonist VPC23019. (A) Cytotoxicity induced
    Inhibitors Jte013, supplied by Tocris, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Biomol GmbH jte 013
    <t>JTE-013,</t> a S1P 2 antagonist, inhibits TNFα induced VCAM-1 and ICAM-1 expression
    Jte 013, supplied by Biomol GmbH, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    JTE-013 decreased the incidence of diabetes. JTE-013 (4 mg/kg) or saline was intraperitoneally injected for 6 days (one shot prior to STZ and five shots with a high dose of STZ (50 mg/kg)). (A) Changes in blood glucose levels of randomly fed mice with/without

    Journal: Biochemical and biophysical research communications

    Article Title: Blockade of sphingosine 1-phosphate receptor 2 signaling attenuates streptozotocin-induced apoptosis of pancreatic ?-cells

    doi: 10.1016/j.bbrc.2010.01.016

    Figure Lengend Snippet: JTE-013 decreased the incidence of diabetes. JTE-013 (4 mg/kg) or saline was intraperitoneally injected for 6 days (one shot prior to STZ and five shots with a high dose of STZ (50 mg/kg)). (A) Changes in blood glucose levels of randomly fed mice with/without

    Article Snippet: JTE-013 (Calbiochem), a specific S1P2 antagonist [ ], was freshly dissolved in saline and intraperitoneally administered at 4 mg/kg for 6 days (one shot prior to STZ and five shots with STZ).

    Techniques: Injection, Mouse Assay

    Stimulation of LPAR 1 /Gi by LPA and S1PR 1/3 /Gi by S1P are critical for stem cell protection. a The expression profiles of LPAR subtypes were determined in cDNAs from hADMSCs (labelled as MSC) or neonatal rat cardiac myocytes for each set of LPAR primers. b Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA treatments, with or without the AM966 (LPAR 1 inhibitor), or PTX (Gi inhibitor) co-treatment ( n = 4). c The expression profiles of S1PR subtypes were determined in cDNAs from hADMSCs (labelled as MSC) for each set of S1PR primers ( n = 4). d Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /S1P treatments, with or without the W146 (S1PR 1 inhibitor), JTE013 (S1PR 2 inhibitor), or CAY10444 (S1PR 3 inhibitor) co-treatment ( n = 4). e Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA/S1P treatments, with or without the PTX (Gi inhibitor) co-treatment ( n = 4). f Knockdown efficiency verification of G 12/13 shRNA transfection using G 12 - and G 13 -specific antibodies. g Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA/S1P treatments, with or without the G 12/13 shRNA co-treatment ( n = 4). h The signalling pathway concluded from this figure. Data from each group are expressed as means ± SEM. Statistical comparison between groups was performed with the Kruskal–Wallis test followed by Dunn’s post-hoc test to detect differences in all groups. *** p

    Journal: Stem Cell Research & Therapy

    Article Title: Co-stimulation of LPAR1 and S1PR1/3 increases the transplantation efficacy of human mesenchymal stem cells in drug-induced and alcoholic liver diseases

    doi: 10.1186/s13287-018-0860-y

    Figure Lengend Snippet: Stimulation of LPAR 1 /Gi by LPA and S1PR 1/3 /Gi by S1P are critical for stem cell protection. a The expression profiles of LPAR subtypes were determined in cDNAs from hADMSCs (labelled as MSC) or neonatal rat cardiac myocytes for each set of LPAR primers. b Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA treatments, with or without the AM966 (LPAR 1 inhibitor), or PTX (Gi inhibitor) co-treatment ( n = 4). c The expression profiles of S1PR subtypes were determined in cDNAs from hADMSCs (labelled as MSC) for each set of S1PR primers ( n = 4). d Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /S1P treatments, with or without the W146 (S1PR 1 inhibitor), JTE013 (S1PR 2 inhibitor), or CAY10444 (S1PR 3 inhibitor) co-treatment ( n = 4). e Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA/S1P treatments, with or without the PTX (Gi inhibitor) co-treatment ( n = 4). f Knockdown efficiency verification of G 12/13 shRNA transfection using G 12 - and G 13 -specific antibodies. g Changes in cell viability and caspase-3/7 activity of hADMSCs after LPS/H 2 O 2 /LPA/S1P treatments, with or without the G 12/13 shRNA co-treatment ( n = 4). h The signalling pathway concluded from this figure. Data from each group are expressed as means ± SEM. Statistical comparison between groups was performed with the Kruskal–Wallis test followed by Dunn’s post-hoc test to detect differences in all groups. *** p

    Article Snippet: All immunophenotypes of the hADMSCs were validated by the manufacturer. d -galactosamine (Gal), lipopolysaccharide (LPS), methylthiazolyldiphenyl tetrazolium bromide (MTT), pertussis toxin (PTX), W146, JTE013, salirasib, and MK-2206 were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Expressing, Activity Assay, shRNA, Transfection

    S1P 2 antagonist JTE-013 inhibited growth of NB xenografts. A, The tumor growth curves of control and JTE-013-treated mice (n=7). B, VEGF mRNA levels in control and JTE-013-treated tumors by quantitative real-time PCR. C , Vessel density in control and

    Journal: Cancer prevention research (Philadelphia, Pa.)

    Article Title: Sphingolipid modulation of angiogenic factor expression in neuroblastoma

    doi: 10.1158/1940-6207.CAPR-11-0017

    Figure Lengend Snippet: S1P 2 antagonist JTE-013 inhibited growth of NB xenografts. A, The tumor growth curves of control and JTE-013-treated mice (n=7). B, VEGF mRNA levels in control and JTE-013-treated tumors by quantitative real-time PCR. C , Vessel density in control and

    Article Snippet: Five days later, the mice were randomized into two groups: JTE-013-treated group receiving 30mg/Kg JTE-013 in 2% (2-hydroxypropyl)-β-cyclodextrin (Sigma) in PBS by gavage daily and the control group receiving the vehicle only.

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction

    Effect of S1PR2 activation on CCA cell migration. Rat BDEsp-TDE H10 cells, human HuCCT1 cells, and CCLP1 cells were plated on six-well plates until confluent. Cells were scratched to simulate a wound and images were recorded as 0 hours. Cells were pretreated with JTE-013 (10 μM) for 1 hour, then treated with TCA (100 μM) or S1P (100 nM) for 48 hours. Images of wound areas were recorded as described in Materials and Methods. The area of wound was quantified using IPLab4.0. Relative wound closure was calculated. (A) Rat BDEsp-TDE H10 cells and human HuCCT1 cells. (B) Human CCLP1 cells. * P

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Conjugated bile acids promote cholangiocarcinoma cell invasive growth through activation of sphingosine 1-phosphate receptor 2

    doi: 10.1002/hep.27085

    Figure Lengend Snippet: Effect of S1PR2 activation on CCA cell migration. Rat BDEsp-TDE H10 cells, human HuCCT1 cells, and CCLP1 cells were plated on six-well plates until confluent. Cells were scratched to simulate a wound and images were recorded as 0 hours. Cells were pretreated with JTE-013 (10 μM) for 1 hour, then treated with TCA (100 μM) or S1P (100 nM) for 48 hours. Images of wound areas were recorded as described in Materials and Methods. The area of wound was quantified using IPLab4.0. Relative wound closure was calculated. (A) Rat BDEsp-TDE H10 cells and human HuCCT1 cells. (B) Human CCLP1 cells. * P

    Article Snippet: Materials S1P and JTE-013 (S1PR2 antagonist) were purchased from Cayman Chemical (Boston, MA).

    Techniques: Activation Assay, Migration

    (A and B) Role of S1PR2 in TCA-mediated cell proliferation in rat BDEsp-TDE H10 cells. (A) Cells were plated in serum-free medium for 24 hours and then treated with TCA (100 μM) with or without JTE-013 (10 μM) for 48 hours. At the end of treatment, viable cells were quantified using the CCK-8 kit, as described in Materials and Methods. ** P

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Conjugated bile acids promote cholangiocarcinoma cell invasive growth through activation of sphingosine 1-phosphate receptor 2

    doi: 10.1002/hep.27085

    Figure Lengend Snippet: (A and B) Role of S1PR2 in TCA-mediated cell proliferation in rat BDEsp-TDE H10 cells. (A) Cells were plated in serum-free medium for 24 hours and then treated with TCA (100 μM) with or without JTE-013 (10 μM) for 48 hours. At the end of treatment, viable cells were quantified using the CCK-8 kit, as described in Materials and Methods. ** P

    Article Snippet: Materials S1P and JTE-013 (S1PR2 antagonist) were purchased from Cayman Chemical (Boston, MA).

    Techniques: CCK-8 Assay

    Effect of TCA and JTE-013 on the expansion of spheroid/“duct-like” structures formed in 3D organotypic cocultures of BDEsp-TDE H10 and BDEsp-TDF E4 cells. Rat BDEsp-TDE H10 and BDEsp-TDF E4 cells were mixed with rat-tail type I collagen gel, as described in Materials and Methods. Cells were treated with TCA (100 μM) or S1P (100 nM) with or without JTE-013 (10 μM) for 8 days. At the end of treatment, the collagen gel cultures were fixed and processed for H E staining. The number and density of spheroid/duct-like structures were quantified as described in Materials and Methods. (A) Representative images of H E staining of spheroid/duct-like structures formed in vehicle control versus S1P or TCA treatment groups with or without JTE-013. (B) The number of spheroid/duct-like structures/cm 2 for each group was quantified as described in Materials and Methods. * P

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Conjugated bile acids promote cholangiocarcinoma cell invasive growth through activation of sphingosine 1-phosphate receptor 2

    doi: 10.1002/hep.27085

    Figure Lengend Snippet: Effect of TCA and JTE-013 on the expansion of spheroid/“duct-like” structures formed in 3D organotypic cocultures of BDEsp-TDE H10 and BDEsp-TDF E4 cells. Rat BDEsp-TDE H10 and BDEsp-TDF E4 cells were mixed with rat-tail type I collagen gel, as described in Materials and Methods. Cells were treated with TCA (100 μM) or S1P (100 nM) with or without JTE-013 (10 μM) for 8 days. At the end of treatment, the collagen gel cultures were fixed and processed for H E staining. The number and density of spheroid/duct-like structures were quantified as described in Materials and Methods. (A) Representative images of H E staining of spheroid/duct-like structures formed in vehicle control versus S1P or TCA treatment groups with or without JTE-013. (B) The number of spheroid/duct-like structures/cm 2 for each group was quantified as described in Materials and Methods. * P

    Article Snippet: Materials S1P and JTE-013 (S1PR2 antagonist) were purchased from Cayman Chemical (Boston, MA).

    Techniques: Staining

    S1P bound to albumin induced PAI-1 expression by activating S1PR2 pathway in 3T3L1 adipocytes. (A) 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM and the medium was exchanged for FBS-free DMEM containing 20 µM of VPC23019, JTE013, or DMSO for 30 minutes. Then, the cells were challenged with a medium containing 10 µM of sphingosine 1-phosphate (S1P) with the corresponding inhibitors for 4 hours. The mRNA level of plasminogen activator inhibitor 1 (PAI-1) was analyzed using real-time PCR. 18S was utilized as an internal control ( n = 4–6/group). † p

    Journal: Journal of Atherosclerosis and Thrombosis

    Article Title: Vehicle-dependent Effects of Sphingosine 1-phosphate on Plasminogen Activator Inhibitor-1 Expression

    doi: 10.5551/jat.37663

    Figure Lengend Snippet: S1P bound to albumin induced PAI-1 expression by activating S1PR2 pathway in 3T3L1 adipocytes. (A) 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM and the medium was exchanged for FBS-free DMEM containing 20 µM of VPC23019, JTE013, or DMSO for 30 minutes. Then, the cells were challenged with a medium containing 10 µM of sphingosine 1-phosphate (S1P) with the corresponding inhibitors for 4 hours. The mRNA level of plasminogen activator inhibitor 1 (PAI-1) was analyzed using real-time PCR. 18S was utilized as an internal control ( n = 4–6/group). † p

    Article Snippet: As shown in , we found that JTE013 inhibited the effect of S1P bound to albumin, while VPC23019 did not inhibit the effect.

    Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction

    S1PR2 mediates Nogo-A-Δ20- and myelin-induced inhibition of cell spreading and neurite outgrowth. (A,C) Representative pictures of 3T3 fibroblasts treated with JTE-013 or vehicle (DMSO) (A), or stably carrying a S1pr2 shRNA (sh- S1pr2 ) or empty vector (sh-Vec) construct (C) and plated on control, Nogo-A-Δ20 or myelin substrates. (B,D) Cell spreading quantification of (A) and (C). (E) Representative pictures of MEFs isolated from WT or S1PR2 −/− mice and plated on control, Nogo-A- Δ20, or myelin substrates. (F) Cell spreading quantification of (E). Cells were stained with Alexa488-conjugated Phalloidin in (A, C, and E). (G,I) Representative pictures of P5–8 cerebellar granule neurons treated with JTE-013 or DMSO (G), or isolated from S1PR2 −/− or WT mice (I) and plated on PLL (ctrl), Nogo-A-Δ20 or myelin substrates. (H,J) Normalized mean neurite length per cell quantification of (G) and (I). Neurons were stained with βIII-Tubulin in (G) and (I). Data shown are means ± SEM ( n = 3–6 experiments; * p

    Journal: PLoS Biology

    Article Title: The Sphingolipid Receptor S1PR2 Is a Receptor for Nogo-A Repressing Synaptic Plasticity

    doi: 10.1371/journal.pbio.1001763

    Figure Lengend Snippet: S1PR2 mediates Nogo-A-Δ20- and myelin-induced inhibition of cell spreading and neurite outgrowth. (A,C) Representative pictures of 3T3 fibroblasts treated with JTE-013 or vehicle (DMSO) (A), or stably carrying a S1pr2 shRNA (sh- S1pr2 ) or empty vector (sh-Vec) construct (C) and plated on control, Nogo-A-Δ20 or myelin substrates. (B,D) Cell spreading quantification of (A) and (C). (E) Representative pictures of MEFs isolated from WT or S1PR2 −/− mice and plated on control, Nogo-A- Δ20, or myelin substrates. (F) Cell spreading quantification of (E). Cells were stained with Alexa488-conjugated Phalloidin in (A, C, and E). (G,I) Representative pictures of P5–8 cerebellar granule neurons treated with JTE-013 or DMSO (G), or isolated from S1PR2 −/− or WT mice (I) and plated on PLL (ctrl), Nogo-A-Δ20 or myelin substrates. (H,J) Normalized mean neurite length per cell quantification of (G) and (I). Neurons were stained with βIII-Tubulin in (G) and (I). Data shown are means ± SEM ( n = 3–6 experiments; * p

    Article Snippet: The slices were pre-incubated for 1 h (or 10 min for the experiments in which JTE-013 and 11c7 were combined) with the inhibitor or DMSO as control in an incubation chamber maintaining a constant flow of the solution.

    Techniques: Inhibition, Stable Transfection, shRNA, Plasmid Preparation, Construct, Isolation, Mouse Assay, Staining

    Nogo-A-Δ20 inhibition is mediated via the G 13 -LARG-RhoA signaling axis and can be modulated by exogenous S1P. (A) 3T3 cells transfected with siRNAs against G 12 , G 13 , G q , or Larg , or control (ctrl) siRNA were replated on a Nogo-A-Δ20 substrate and assessed for cell spreading. G i/o was blocked with Pertussis Toxin (PTX) for which saline was used as control. JTE-013 was co-applied to G 13 -siRNA-treated cells to investigate a cumulative effect. (B) Transfection of DIV4 E19 cortical neurons with siRNA against G 13 but not G 12 similarly rescued Nogo-A-Δ20-induced neurite outgrowth inhibition. (C,D) Nogo-A-Δ20-induced RhoA activation was assessed in JTE-013- versus DMSO-treated cells (C) or in cells carrying a stable knockdown of S1PR2 (sh- S1pr2 ) versus control vector (sh-Vec) (D). (E,F) Relative quantification of (C) and (D), respectively. (G,H) Competitive ELISA quantifications of extra- (EC) and intracellular (IC) S1P levels in 3T3 cells (G) and cerebellar granule neurons (H) before and after 30 and 60 min incubation with Nogo-A-Δ20. (I) Quantification of Nogo-A-Δ20-mediated cell spreading inhibition in the presence of the SphK-specific blocker D,L- threo -dihydrosphingosine (DHS) or in SphK1 −/− or SphK2 −/− MEFs. (J,K) 3T3 cells were plated on a Nogo-A-Δ20 substrate in the presence of the function blocking anti-S1P antibody Sphingomab (J) or of exogenous S1P (K) and assessed for cell spreading. Co-application of JTE-013 significantly reversed the modulatory effects obtained by S1P (K) but not anti-S1P (J). Anti-BrdU antibody or methanol was used as control in (J) and (K). Data shown are means ± SEM ( n = 3–6 experiments; * p

    Journal: PLoS Biology

    Article Title: The Sphingolipid Receptor S1PR2 Is a Receptor for Nogo-A Repressing Synaptic Plasticity

    doi: 10.1371/journal.pbio.1001763

    Figure Lengend Snippet: Nogo-A-Δ20 inhibition is mediated via the G 13 -LARG-RhoA signaling axis and can be modulated by exogenous S1P. (A) 3T3 cells transfected with siRNAs against G 12 , G 13 , G q , or Larg , or control (ctrl) siRNA were replated on a Nogo-A-Δ20 substrate and assessed for cell spreading. G i/o was blocked with Pertussis Toxin (PTX) for which saline was used as control. JTE-013 was co-applied to G 13 -siRNA-treated cells to investigate a cumulative effect. (B) Transfection of DIV4 E19 cortical neurons with siRNA against G 13 but not G 12 similarly rescued Nogo-A-Δ20-induced neurite outgrowth inhibition. (C,D) Nogo-A-Δ20-induced RhoA activation was assessed in JTE-013- versus DMSO-treated cells (C) or in cells carrying a stable knockdown of S1PR2 (sh- S1pr2 ) versus control vector (sh-Vec) (D). (E,F) Relative quantification of (C) and (D), respectively. (G,H) Competitive ELISA quantifications of extra- (EC) and intracellular (IC) S1P levels in 3T3 cells (G) and cerebellar granule neurons (H) before and after 30 and 60 min incubation with Nogo-A-Δ20. (I) Quantification of Nogo-A-Δ20-mediated cell spreading inhibition in the presence of the SphK-specific blocker D,L- threo -dihydrosphingosine (DHS) or in SphK1 −/− or SphK2 −/− MEFs. (J,K) 3T3 cells were plated on a Nogo-A-Δ20 substrate in the presence of the function blocking anti-S1P antibody Sphingomab (J) or of exogenous S1P (K) and assessed for cell spreading. Co-application of JTE-013 significantly reversed the modulatory effects obtained by S1P (K) but not anti-S1P (J). Anti-BrdU antibody or methanol was used as control in (J) and (K). Data shown are means ± SEM ( n = 3–6 experiments; * p

    Article Snippet: The slices were pre-incubated for 1 h (or 10 min for the experiments in which JTE-013 and 11c7 were combined) with the inhibitor or DMSO as control in an incubation chamber maintaining a constant flow of the solution.

    Techniques: Inhibition, Transfection, Activation Assay, Plasmid Preparation, Competitive ELISA, Incubation, Blocking Assay

    Blockade of S1PR2 phenocopies the increase in hippocampal and cortical LTP observed upon Nogo-A neutralization. (A,B) Hippocampal WT (A) and Nogo-A −/− (B) slices were treated with JTE-013 or vehicle (DMSO) (WT DMSO : n = 8; Nogo-A −/− DMSO : n = 10; WT JTE-013 : n = 11; Nogo-A −/− JTE-013 : n = 9). 60 min after theta-burst stimulation (arrow), a significant difference in LTP could be observed between JTE-013 and DMSO treatment in WT (A) but not Nogo-A −/− (B) slices. (C,D) Input-output strength revealed no differences in JTE-013- versus DMSO-treated slices of WT (C) and Nogo-A −/− (D) mice (WT DMSO : n = 6; Nogo-A −/− DMSO : n = 6; WT JTE-013 : n = 7; Nogo-A −/− JTE-013 : n = 6). (E,F) PPF revealed no alterations in JTE-013- versus DMSO-treated slices of WT (E) and Nogo-A −/− (F) mice (WT DMSO : n = 7; Nogo-A −/− DMSO : n = 6; WT JTE-013 : n = 5; Nogo-A −/− JTE-013 : n = 6). (G) LTP was measured upon simultaneous neutralization of S1PR2 using JTE-013 and of Nogo-A using 11c7 (IgG1 + DMSO: n = 7; IgG1 + JTE-013: n = 6; 11c7 + DMSO: n = 8; 11c7 + JTE-013: n = 6). (H) LTP was measured upon simultaneous neutralization of S1PR2 using JTE-013 and of NgR1 using anti-NgR1 (DMSO: n = 7; JTE-013: n = 9; anti-NgR1 + JTE-013: n = 8). (I) Rat motor forelimb area brain slices were treated with JTE-013 ( n = 7) or DMSO ( n = 8). Peak amplitudes were significantly larger in JTE-013- versus DMSO-treated slices upon repeated inductions of LTP (multiple arrows). (J) Input-output strength revealed no differences in JTE-013- ( n = 8) versus DMSO-treated ( n = 12) cortical slices. Insets show representative traces. Data shown are means ± SEM (* p

    Journal: PLoS Biology

    Article Title: The Sphingolipid Receptor S1PR2 Is a Receptor for Nogo-A Repressing Synaptic Plasticity

    doi: 10.1371/journal.pbio.1001763

    Figure Lengend Snippet: Blockade of S1PR2 phenocopies the increase in hippocampal and cortical LTP observed upon Nogo-A neutralization. (A,B) Hippocampal WT (A) and Nogo-A −/− (B) slices were treated with JTE-013 or vehicle (DMSO) (WT DMSO : n = 8; Nogo-A −/− DMSO : n = 10; WT JTE-013 : n = 11; Nogo-A −/− JTE-013 : n = 9). 60 min after theta-burst stimulation (arrow), a significant difference in LTP could be observed between JTE-013 and DMSO treatment in WT (A) but not Nogo-A −/− (B) slices. (C,D) Input-output strength revealed no differences in JTE-013- versus DMSO-treated slices of WT (C) and Nogo-A −/− (D) mice (WT DMSO : n = 6; Nogo-A −/− DMSO : n = 6; WT JTE-013 : n = 7; Nogo-A −/− JTE-013 : n = 6). (E,F) PPF revealed no alterations in JTE-013- versus DMSO-treated slices of WT (E) and Nogo-A −/− (F) mice (WT DMSO : n = 7; Nogo-A −/− DMSO : n = 6; WT JTE-013 : n = 5; Nogo-A −/− JTE-013 : n = 6). (G) LTP was measured upon simultaneous neutralization of S1PR2 using JTE-013 and of Nogo-A using 11c7 (IgG1 + DMSO: n = 7; IgG1 + JTE-013: n = 6; 11c7 + DMSO: n = 8; 11c7 + JTE-013: n = 6). (H) LTP was measured upon simultaneous neutralization of S1PR2 using JTE-013 and of NgR1 using anti-NgR1 (DMSO: n = 7; JTE-013: n = 9; anti-NgR1 + JTE-013: n = 8). (I) Rat motor forelimb area brain slices were treated with JTE-013 ( n = 7) or DMSO ( n = 8). Peak amplitudes were significantly larger in JTE-013- versus DMSO-treated slices upon repeated inductions of LTP (multiple arrows). (J) Input-output strength revealed no differences in JTE-013- ( n = 8) versus DMSO-treated ( n = 12) cortical slices. Insets show representative traces. Data shown are means ± SEM (* p

    Article Snippet: The slices were pre-incubated for 1 h (or 10 min for the experiments in which JTE-013 and 11c7 were combined) with the inhibitor or DMSO as control in an incubation chamber maintaining a constant flow of the solution.

    Techniques: Neutralization, Mouse Assay

    AC/S1P promote Akt-dependent export of nuclear PTEN. PPC1 cells were transfected with WT-PTEN were infected with Ad-GFP or Ad-AC for 48 hours in the presence of DMSO (NT) or the indicated compounds for 24 hours. A) Nuclear fractions from the indicated treatments were isolated and evaluated for presence of PTEN with Histone H3 as a nuclear loading control and absence of β-tubulin to indicate purity of the nuclear sample. B) Cells were immunostained for PTEN (red) and nuclei (blue). C) The percentage of cells from (B) which had nuclear PTEN in each treatment. D) PPC1 cells transfected with WT-PTEN were treated with 1µM JTE013 or 5µM AktX for 24 hours prior to treatment with the indicated dose of S1P or PBS for 2 hours followed by fixation and immunostaining for PTEN (red) and nuclei (blue). E) The percentage of cells from (D) which had nuclear PTEN. F) Nuclear fractions from the indicated treatments were isolated and evaluated for presence of PTEN with Histone H3 as a nuclear loading control and absence of β-tubulin to indicate purity of the nuclear sample. One way ANOVA with Bonferroni correction, *p

    Journal: PLoS ONE

    Article Title: Acid Ceramidase Promotes Nuclear Export of PTEN through Sphingosine 1-Phosphate Mediated Akt Signaling

    doi: 10.1371/journal.pone.0076593

    Figure Lengend Snippet: AC/S1P promote Akt-dependent export of nuclear PTEN. PPC1 cells were transfected with WT-PTEN were infected with Ad-GFP or Ad-AC for 48 hours in the presence of DMSO (NT) or the indicated compounds for 24 hours. A) Nuclear fractions from the indicated treatments were isolated and evaluated for presence of PTEN with Histone H3 as a nuclear loading control and absence of β-tubulin to indicate purity of the nuclear sample. B) Cells were immunostained for PTEN (red) and nuclei (blue). C) The percentage of cells from (B) which had nuclear PTEN in each treatment. D) PPC1 cells transfected with WT-PTEN were treated with 1µM JTE013 or 5µM AktX for 24 hours prior to treatment with the indicated dose of S1P or PBS for 2 hours followed by fixation and immunostaining for PTEN (red) and nuclei (blue). E) The percentage of cells from (D) which had nuclear PTEN. F) Nuclear fractions from the indicated treatments were isolated and evaluated for presence of PTEN with Histone H3 as a nuclear loading control and absence of β-tubulin to indicate purity of the nuclear sample. One way ANOVA with Bonferroni correction, *p

    Article Snippet: DU145 cells were treated with A) 1µM JTE013 or DMSO (NT) or B) 5µM AktX or water (NT) for 24 hours prior to stimulation with 500 nM S1P or PBS (NT) for 2 hours.

    Techniques: Transfection, Infection, Isolation, Immunostaining

    Receptor-selective kinase pathway activation. hMSC-TERT cells in base media were pretreated for 15 min with either DMSO (vehicle) or the indicated inhibitor and a S1P receptor antagonist (receptor antagonist to S1PR2 (JTE-013), thus measuring the impacts

    Journal: The Journal of Biological Chemistry

    Article Title: Sphingosine 1-Phosphate (S1P) Receptors 1 and 2 Coordinately Induce Mesenchymal Cell Migration through S1P Activation of Complementary Kinase Pathways *

    doi: 10.1074/jbc.M112.413583

    Figure Lengend Snippet: Receptor-selective kinase pathway activation. hMSC-TERT cells in base media were pretreated for 15 min with either DMSO (vehicle) or the indicated inhibitor and a S1P receptor antagonist (receptor antagonist to S1PR2 (JTE-013), thus measuring the impacts

    Article Snippet: In some experiments vehicle (5% acidified DMSO in H2 O), combined S1PR1,3 (VPC23019, 100 n m , Avanti Polar Lipids), and S1PR2 (JTE-013, 20 n m , Tocris Bioscience) antagonism or selective receptor antagonists were employed.

    Techniques: Activation Assay

    Schematic of coupling and S1P signaling in mesenchymal cells. Osteoclast SPHK generates S1P, which activates receptors S1PR1 and S1PR2 on mesenchymal cells. S1PR1 activated the JAK/STAT pathway, and S1PR2 activates the FAK/PI3K/AKT pathway to stimulate

    Journal: The Journal of Biological Chemistry

    Article Title: Sphingosine 1-Phosphate (S1P) Receptors 1 and 2 Coordinately Induce Mesenchymal Cell Migration through S1P Activation of Complementary Kinase Pathways *

    doi: 10.1074/jbc.M112.413583

    Figure Lengend Snippet: Schematic of coupling and S1P signaling in mesenchymal cells. Osteoclast SPHK generates S1P, which activates receptors S1PR1 and S1PR2 on mesenchymal cells. S1PR1 activated the JAK/STAT pathway, and S1PR2 activates the FAK/PI3K/AKT pathway to stimulate

    Article Snippet: In some experiments vehicle (5% acidified DMSO in H2 O), combined S1PR1,3 (VPC23019, 100 n m , Avanti Polar Lipids), and S1PR2 (JTE-013, 20 n m , Tocris Bioscience) antagonism or selective receptor antagonists were employed.

    Techniques:

    Rho GTPase activation. A , hMSC-TERT cells were treated with either BASE with vehicle or S1P agonist or osteoclast conditioned medium ( OC CM ) combined with either vehicle or a S1P receptor antagonists (S1PR1 and S1PR2 antagonists together) (W123 and JTE-013)

    Journal: The Journal of Biological Chemistry

    Article Title: Sphingosine 1-Phosphate (S1P) Receptors 1 and 2 Coordinately Induce Mesenchymal Cell Migration through S1P Activation of Complementary Kinase Pathways *

    doi: 10.1074/jbc.M112.413583

    Figure Lengend Snippet: Rho GTPase activation. A , hMSC-TERT cells were treated with either BASE with vehicle or S1P agonist or osteoclast conditioned medium ( OC CM ) combined with either vehicle or a S1P receptor antagonists (S1PR1 and S1PR2 antagonists together) (W123 and JTE-013)

    Article Snippet: In some experiments vehicle (5% acidified DMSO in H2 O), combined S1PR1,3 (VPC23019, 100 n m , Avanti Polar Lipids), and S1PR2 (JTE-013, 20 n m , Tocris Bioscience) antagonism or selective receptor antagonists were employed.

    Techniques: Activation Assay

    The relationship between Foxo3a and S1PR2 in PBMCs (A) and (B) PBMCs were pre-treated with LY294002 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α I and IFN γ were detected by RT-PCR. (C) and (D) PBMCs were pre-treated with SC79 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. (E) and (F) PBMCs were treated with LPS and JET-013 for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. *, p

    Journal: Oncotarget

    Article Title: Changes of Foxo3a in PBMCs and its associations with stress hyperglycemia in acute obstructive suppurative cholangitis patients

    doi: 10.18632/oncotarget.20011

    Figure Lengend Snippet: The relationship between Foxo3a and S1PR2 in PBMCs (A) and (B) PBMCs were pre-treated with LY294002 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α I and IFN γ were detected by RT-PCR. (C) and (D) PBMCs were pre-treated with SC79 for 1 h before stimulation with LPS for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. (E) and (F) PBMCs were treated with LPS and JET-013 for 2 h: the protein and phosphorylation levels of Foxo3a and S1PR2 were tested by WB assay; the mRNA levels of TNF α and IFN γ were detected by RT-PCR. *, p

    Article Snippet: The adherent cells continued to cultivate for 12 h. Then, PBMCs were pretreated with LY294002 (30 μM) (S1105, Selleck, USA) and SC79 (4 μg/ml) (S7863, Selleck, USA) for 1 h, followed by treatment with LPS (100 ng/ml) (L5293, SIGMA, USA) for 2 h. For S1PR2 inhibitor testing, PBMCs were pretreated with JET-013 (1 μM) (S7182, Selleck, the USA) and LPS (100 ng/ml) for 2 h. The total protein and RNA of PBMCs were collected for western blotting (WB) and real-time polymerase chain reaction (RT-PCR).

    Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction

    Activation of NF-κB and PI3K/Akt pathways in PBMCs (A) The expression and phosphorylation levels of Foxo3a were tested by WB in two patients before (AP) and after (RP) treatment and in one healthy volunteer (HV). (B) The protein levels of p-IκB-α and p-NF-κB p65 increased dramatically in the AP in AOSC patients and decreased to normal levels after treatment, while the expression of p-PI3K, p-Akt and S1PR2 dramatically decreased after treatment. *, p

    Journal: Oncotarget

    Article Title: Changes of Foxo3a in PBMCs and its associations with stress hyperglycemia in acute obstructive suppurative cholangitis patients

    doi: 10.18632/oncotarget.20011

    Figure Lengend Snippet: Activation of NF-κB and PI3K/Akt pathways in PBMCs (A) The expression and phosphorylation levels of Foxo3a were tested by WB in two patients before (AP) and after (RP) treatment and in one healthy volunteer (HV). (B) The protein levels of p-IκB-α and p-NF-κB p65 increased dramatically in the AP in AOSC patients and decreased to normal levels after treatment, while the expression of p-PI3K, p-Akt and S1PR2 dramatically decreased after treatment. *, p

    Article Snippet: The adherent cells continued to cultivate for 12 h. Then, PBMCs were pretreated with LY294002 (30 μM) (S1105, Selleck, USA) and SC79 (4 μg/ml) (S7863, Selleck, USA) for 1 h, followed by treatment with LPS (100 ng/ml) (L5293, SIGMA, USA) for 2 h. For S1PR2 inhibitor testing, PBMCs were pretreated with JET-013 (1 μM) (S7182, Selleck, the USA) and LPS (100 ng/ml) for 2 h. The total protein and RNA of PBMCs were collected for western blotting (WB) and real-time polymerase chain reaction (RT-PCR).

    Techniques: Activation Assay, Expressing, Western Blot

    Rho GTPase activation. A , hMSC-TERT cells were treated with either BASE with vehicle or S1P agonist or osteoclast conditioned medium ( OC CM ) combined with either vehicle or a S1P receptor antagonists (S1PR1 and S1PR2 antagonists together) (W123 and JTE-013)

    Journal: The Journal of Biological Chemistry

    Article Title: Sphingosine 1-Phosphate (S1P) Receptors 1 and 2 Coordinately Induce Mesenchymal Cell Migration through S1P Activation of Complementary Kinase Pathways *

    doi: 10.1074/jbc.M112.413583

    Figure Lengend Snippet: Rho GTPase activation. A , hMSC-TERT cells were treated with either BASE with vehicle or S1P agonist or osteoclast conditioned medium ( OC CM ) combined with either vehicle or a S1P receptor antagonists (S1PR1 and S1PR2 antagonists together) (W123 and JTE-013)

    Article Snippet: The selective antagonists were S1PR1 antagonist W123, 1 μ m , Cayman Chemical; S1PR2 antagonist JTE-013, 20 n m ; S1PR3 antagonist BML-241, 10 μ m, Cayman Chemical.

    Techniques: Activation Assay

    Effect of CM from MDA-MB-231 cells on ERK-1/2 activation and DNA synthesis in MEFs MEFs quiescent for 24 hr were treated with non-conditioned medium (NCM) or conditioned medium (CM) from MDA-MB-231 cells for 10 min. MEFs were pre-treated with and without JTE-013 (10 μM) for 15 min prior to addition of NCM or CM. MEFs were also treated with CYM5520 (10 μM) in the DNA synthesis experiments. CM was also isolated from MDA-MB-231 cells treated with scrambled or S1P 2 siRNA for 24 h. (A) Western blot showing the effect of CM from MDA-MB-231 cells and JTE-013 on ERK-1/2 activation in MEFs. (B) Bar graph showing the effect of NCM, CM from MDA-MB-231 cells or CYM5520 on [ 3 H]-thymidine incorporation into DNA synthesis in MEFs. Results are expressed as means +/- SEM for n=3. (C) Western blot showing the effect of CM on ERK-1/2 activation in MEFs. CM was isolated from MDA-MB-231 cells that had been treated with either scrambled siRNA (200 nM) or S1P 2 siRNA (100 or 200 nM). Also shown is the effect of S1P 2 siRNA on S1P 2 expression levels in MDA-MB-231 cells. (D) Western blot showing that neither P-ERK-1/2 nor ERK-2 is released from MDA-MB-231 cells into CM as evidenced by their absence in the PPT. (E) Western blot showing the lack of effect of CM from MDA-MB-453 cells on ERK-1/2 activation in MEFs. Blots were probed with anti-phospho ERK-1/2 and anti-S1P 2 antibodies. (A, C, D, E). Blots were re-probed with anti-actin antibody to ensure equal protein loading (A, C, E). Results (A, C, D, E) are representative of 3 independent experiments.

    Journal: Oncotarget

    Article Title: The sphingosine 1-phosphate receptor 2 is shed in exosomes from breast cancer cells and is N-terminally processed to a short constitutively active form that promotes extracellular signal regulated kinase activation and DNA synthesis in fibroblasts

    doi: 10.18632/oncotarget.25658

    Figure Lengend Snippet: Effect of CM from MDA-MB-231 cells on ERK-1/2 activation and DNA synthesis in MEFs MEFs quiescent for 24 hr were treated with non-conditioned medium (NCM) or conditioned medium (CM) from MDA-MB-231 cells for 10 min. MEFs were pre-treated with and without JTE-013 (10 μM) for 15 min prior to addition of NCM or CM. MEFs were also treated with CYM5520 (10 μM) in the DNA synthesis experiments. CM was also isolated from MDA-MB-231 cells treated with scrambled or S1P 2 siRNA for 24 h. (A) Western blot showing the effect of CM from MDA-MB-231 cells and JTE-013 on ERK-1/2 activation in MEFs. (B) Bar graph showing the effect of NCM, CM from MDA-MB-231 cells or CYM5520 on [ 3 H]-thymidine incorporation into DNA synthesis in MEFs. Results are expressed as means +/- SEM for n=3. (C) Western blot showing the effect of CM on ERK-1/2 activation in MEFs. CM was isolated from MDA-MB-231 cells that had been treated with either scrambled siRNA (200 nM) or S1P 2 siRNA (100 or 200 nM). Also shown is the effect of S1P 2 siRNA on S1P 2 expression levels in MDA-MB-231 cells. (D) Western blot showing that neither P-ERK-1/2 nor ERK-2 is released from MDA-MB-231 cells into CM as evidenced by their absence in the PPT. (E) Western blot showing the lack of effect of CM from MDA-MB-453 cells on ERK-1/2 activation in MEFs. Blots were probed with anti-phospho ERK-1/2 and anti-S1P 2 antibodies. (A, C, D, E). Blots were re-probed with anti-actin antibody to ensure equal protein loading (A, C, E). Results (A, C, D, E) are representative of 3 independent experiments.

    Article Snippet: CYM5520 and JTE-013 were purchased from Cambridge Biosciences (Cambridge, UK).

    Techniques: Multiple Displacement Amplification, Activation Assay, DNA Synthesis, Isolation, Western Blot, Expressing

    Uptake of exosomal S1P 2 from breast cancer cells and processing by MEFs MEFs quiescent for 24 hr were treated with NCM or CM from MDA-MB-231 cells for 10 min. MEFs were pre-treated with and without JTE-013 (10 μM) or CYM5520 (10, 25 μM) for 15 min prior to addition of NCM or CM. (A) Western blot showing the lack of effect of CYM5520 alone on ERK-1/2 activation in MEFs. (B) Western blot showing the uptake into MEFs of the short S1P 2 (Mr = 36 kDa) form and the activation of ERK-1/2 in response to CM isolated from MDA-MB-231 cells. (A) and (B) Blots were re-probed with anti-actin antibody to ensure equal protein loading. (C) Immunofluorescence images of control, CM-stimulated and CYM5520 (10 μM)-stimulated MEFs co-stained with anti-actin (TRITC secondary antibody, red) and anti-S1P 2 (FITC secondary antibody, green) antibodies. (D) Fluorescence image of MEFs showing uptake of GFP-hsp70 by detection of GFP and co-stained with anti-actin antibody using a Texas Red conjugated secondary antibody or DAPI. Results are representative of 3 independent experiments.

    Journal: Oncotarget

    Article Title: The sphingosine 1-phosphate receptor 2 is shed in exosomes from breast cancer cells and is N-terminally processed to a short constitutively active form that promotes extracellular signal regulated kinase activation and DNA synthesis in fibroblasts

    doi: 10.18632/oncotarget.25658

    Figure Lengend Snippet: Uptake of exosomal S1P 2 from breast cancer cells and processing by MEFs MEFs quiescent for 24 hr were treated with NCM or CM from MDA-MB-231 cells for 10 min. MEFs were pre-treated with and without JTE-013 (10 μM) or CYM5520 (10, 25 μM) for 15 min prior to addition of NCM or CM. (A) Western blot showing the lack of effect of CYM5520 alone on ERK-1/2 activation in MEFs. (B) Western blot showing the uptake into MEFs of the short S1P 2 (Mr = 36 kDa) form and the activation of ERK-1/2 in response to CM isolated from MDA-MB-231 cells. (A) and (B) Blots were re-probed with anti-actin antibody to ensure equal protein loading. (C) Immunofluorescence images of control, CM-stimulated and CYM5520 (10 μM)-stimulated MEFs co-stained with anti-actin (TRITC secondary antibody, red) and anti-S1P 2 (FITC secondary antibody, green) antibodies. (D) Fluorescence image of MEFs showing uptake of GFP-hsp70 by detection of GFP and co-stained with anti-actin antibody using a Texas Red conjugated secondary antibody or DAPI. Results are representative of 3 independent experiments.

    Article Snippet: CYM5520 and JTE-013 were purchased from Cambridge Biosciences (Cambridge, UK).

    Techniques: Multiple Displacement Amplification, Western Blot, Activation Assay, Isolation, Immunofluorescence, Staining, Fluorescence

    Effect of S1PR1-3 inhibition on angiogenesis in vitro (A) Representative images of the migration, invasion and tube formation assays. Endothelial cells were stimulated with CM collected from the ovarian cancer cells precultured with VPC23019 (300nM) or JTE-013 (1μM). Migrated cells, invaded cells and tube like structures were photographed. (B) Statistical analysis of the migrated cells, invaded cells and tube like structures. (C) Effect of VPC23019 and JTE-013 on the VEGF, IL-8 and IL-6 secretion of ovarian cancer cells. All experiments were repeated three times, with three replicates in each group ( * p

    Journal: Oncotarget

    Article Title: Sphingosine kinase 1/sphingosine-1-phosphate (S1P)/S1P receptor axis is involved in ovarian cancer angiogenesis

    doi: 10.18632/oncotarget.20471

    Figure Lengend Snippet: Effect of S1PR1-3 inhibition on angiogenesis in vitro (A) Representative images of the migration, invasion and tube formation assays. Endothelial cells were stimulated with CM collected from the ovarian cancer cells precultured with VPC23019 (300nM) or JTE-013 (1μM). Migrated cells, invaded cells and tube like structures were photographed. (B) Statistical analysis of the migrated cells, invaded cells and tube like structures. (C) Effect of VPC23019 and JTE-013 on the VEGF, IL-8 and IL-6 secretion of ovarian cancer cells. All experiments were repeated three times, with three replicates in each group ( * p

    Article Snippet: VPC23019 and JTE-013 was purchased from Santa Cruz Biotechnology.

    Techniques: Inhibition, In Vitro, Migration

    S1PR4 on DC conveys ACM-dependent suppression of cytotoxicity. (A–D) T cells were co-cultured with control or ACM-primed autologous DC with or without the S1PR2/4 antagonist JTE-013 or the S1PR1/3 antagonist VPC23019. (A) Cytotoxicity induced

    Journal: European journal of immunology

    Article Title: Apoptotic tumor cells induce IL-27 release from human DC to activate regulatory T cells that express CD69 and attenuate cytotoxicity

    doi: 10.1002/eji.201142093

    Figure Lengend Snippet: S1PR4 on DC conveys ACM-dependent suppression of cytotoxicity. (A–D) T cells were co-cultured with control or ACM-primed autologous DC with or without the S1PR2/4 antagonist JTE-013 or the S1PR1/3 antagonist VPC23019. (A) Cytotoxicity induced

    Article Snippet: S1PR2/4 antagonist JTE-013 (15 μM) (Biomol, Hamburg Germany) and the S1PR4 antagonists and (each 200 nM) [ ] were dissolved in DMSO.

    Techniques: Cell Culture

    JTE-013, a S1P 2 antagonist, inhibits TNFα induced VCAM-1 and ICAM-1 expression

    Journal: Prostaglandins & other lipid mediators

    Article Title: Sphingosine-1-phosphate receptor-2 mediated NFκB activation contributes to tumor necrosis factor-α induced VCAM-1 and ICAM-1 expression in endothelial cells

    doi: 10.1016/j.prostaglandins.2013.06.001

    Figure Lengend Snippet: JTE-013, a S1P 2 antagonist, inhibits TNFα induced VCAM-1 and ICAM-1 expression

    Article Snippet: Sphingosine-1-phosphate was purchased from Biomol; JTE-013 and VPC23019 were from Avanti.

    Techniques: Expressing