anti s1p1  (Alomone Labs)

Journal: Frontiers in Psychiatry

Article Title: Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2

doi: 10.3389/fpsyt.2022.827981

Figure Lengend Snippet: Representative images of [ 3 H]CS1P1 autoradiograph, S1PR1 immunostaining, and Hematoxylin and eosin (H E) staining in postmortem human DLPFC tissues. The distribution of [ 3 H]CS1P1 matched well with anti-S1PR1 antibody, and was mainly located in the gray matter regions as indicated in the H E staining.

Article Snippet: After that, all sections were stained with anti-S1PR1 antibody (Alomone, Jerusalem, Israel) overnight at 4°C, washed and followed by incubation with ImmPRESS HRP Horse anti-rabbit polymer for 1 h at RT, and developed using ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Autoradiography, Immunostaining, Staining

Journal: Frontiers in Psychiatry

Article Title: Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2

doi: 10.3389/fpsyt.2022.827981

Figure Lengend Snippet: ARG S1PR1 intensity expression (in fmol/mg) comparison between normal controls, schizophrenia Type 1, and schizophrenia Type 2 (* p

Article Snippet: After that, all sections were stained with anti-S1PR1 antibody (Alomone, Jerusalem, Israel) overnight at 4°C, washed and followed by incubation with ImmPRESS HRP Horse anti-rabbit polymer for 1 h at RT, and developed using ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Expressing

Journal: Frontiers in Psychiatry

Article Title: Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2

doi: 10.3389/fpsyt.2022.827981

Figure Lengend Snippet: Immunohistochemistry (IHC) of S1PR1 in postmortem DLPFC tissues from the representative normal control and schizophrenia Type 1 and Type 2.

Article Snippet: After that, all sections were stained with anti-S1PR1 antibody (Alomone, Jerusalem, Israel) overnight at 4°C, washed and followed by incubation with ImmPRESS HRP Horse anti-rabbit polymer for 1 h at RT, and developed using ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Immunohistochemistry

Journal: Frontiers in Psychiatry

Article Title: Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2

doi: 10.3389/fpsyt.2022.827981

Figure Lengend Snippet: Autoradiography images of S1PR1 using [ 3 H]CS1P1 in postmortem DLPFC tissues from representative normal control, schizophrenia Type 1, and schizophrenia Type 2. In general, [ 3 H]CS1P1 was higher in Type 2 schizophrenia subjects compared with normal control and Type 1 schizophrenia subjects.

Article Snippet: After that, all sections were stained with anti-S1PR1 antibody (Alomone, Jerusalem, Israel) overnight at 4°C, washed and followed by incubation with ImmPRESS HRP Horse anti-rabbit polymer for 1 h at RT, and developed using ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Autoradiography

Journal: Frontiers in Psychiatry

Article Title: Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2

doi: 10.3389/fpsyt.2022.827981

Figure Lengend Snippet: ARG S1PR1 intensity expression (fmol/mg) triplicate measures (M1, M2, and M3) in the DLPFC from normal controls, schizophrenia Type 1, and schizophrenia Type 2.

Article Snippet: After that, all sections were stained with anti-S1PR1 antibody (Alomone, Jerusalem, Israel) overnight at 4°C, washed and followed by incubation with ImmPRESS HRP Horse anti-rabbit polymer for 1 h at RT, and developed using ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Expressing

Journal: Neural Plasticity

Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System

doi: 10.1155/2017/6818970

Figure Lengend Snippet: Modulation of S1PR1 expression by the CNTF/Stat3 pathway in optic nerve-injured retinae. (a, b) S1PR1 expression changes were monitored 3 d after ONC in retinae infected with ShH10.CNTF or ShH10.Empty, a control vector that was deprived of cDNA sequence. ShH10 viruses preferentially infected Müller glia in the retina [ 16 ]. P-Stat3 and S1PR1 were markedly increased by ShH10.CNTF in RGC somata identified using β 3-tubulin as a specific marker. (c) Five days after ONC, qRT-PCR measurements showed that the infection of RGCs with AAV2.Stat3 significantly increased the mRNA level of S1pr1 compared with control AAV2.GFP vector. ShH10 and AAV2 viruses were intravitreally injected 4 weeks before ONC. Three mice were analyzed/grouped. Statistics: one-way ANOVA; ∗ p

Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).

Techniques: Expressing, Infection, Plasmid Preparation, Sequencing, Marker, Quantitative RT-PCR, Injection, Mouse Assay

Journal: Neural Plasticity

Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System

doi: 10.1155/2017/6818970

Figure Lengend Snippet: S1PR1 knockdown alters CNTF-induced RGC survival after ONC. (a) Two weeks after ONC, surviving RGCs were observed in retinal flat-mounts after immunofluorescent staining for β 3-tubulin. Less RGCs were visible in retinae infected with AAV2.shRNA-S1PR1 and ShH10.CNTF ( n = 5 mice) than in mice injected with ShH10.CNTF/AAV2.GFP ( n = 7 mice) or ShH10.CNTF/AAV2.shRNA-S1PR1 ( n = 5 mice). (b) Quantitatively, the average number of surviving RGCs was statistically lower in whole retinae transduced with ShH10.CNTF/AAV2.shRNA-S1PR1 than in the two other groups of animals (ANOVA, ∗∗∗ p

Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).

Techniques: Staining, Infection, shRNA, Mouse Assay, Injection, Transduction

Journal: Neural Plasticity

Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System

doi: 10.1155/2017/6818970

Figure Lengend Snippet: The expression of P-Stat3 is not changed by S1PR1 silencing after CNTF stimulation. The expression of P-Stat3 was assessed by western blotting in protein lysates (20 μ g) from retinae treated with ShH10 and AAV2 viruses. P-Stat3 and Stat3 blots were quantified by densitometry using the ImageJ software (NIH). The level of P-Stat3/Stat3 was not significantly different between mice treated with ShH10.CNTF/AAV2.shRNA-S1PR1 and ShH10.CNTF/AAV2.GFP. Three mice were analyzed for each group.

Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).

Techniques: Expressing, Western Blot, Software, Mouse Assay, shRNA

Journal: Neural Plasticity

Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System

doi: 10.1155/2017/6818970

Figure Lengend Snippet: Hypothetical mechanism by which CNTF/Stat3 and S1P/S1PR1 interaction may orchestrate neuronal survival and axonal growth. CNTF binds and activates a heterotrimeric receptor complex, composed of CNTFR α , leukemia inhibitory factor receptor (LIFR), and gp130, leading to Stat3 phosphorylation (P-Stat3) and activation. (a) P-Stat3-driven transcription may increase the expression of S1PR1 and its translocation to the plasma membrane. The activation of S1PR1 by S1P may trigger downstream growth mechanisms resulting in (b) neuronal survival and (c) axonal growth.

Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).

Techniques: Activation Assay, Expressing, Translocation Assay

Journal: Neural Plasticity

Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System

doi: 10.1155/2017/6818970

Figure Lengend Snippet: S1PR1 knockdown potentiates CNTF-induced axonal regeneration. (a) Axonal regeneration was visualized on longitudinal sections of optic nerves two weeks after crush injury and 4 weeks after coinfection with ShH10.CNTF and AAV2 vectors. Axons were traced with cholera toxin β subunit (CTb) conjugated to Alexa 594 the day before tissue fixation. (b) The infection of retinal cells with ShH10.CNTF and AAV2.shRNA-S1PR1 promoted lengthy axonal regeneration in the optic nerve compared with the ShH10.CNTF/AAV2.GFP combination. (c) Quantitatively, axonal fibers were significantly more numerous between 1300 and 1800 μ m past the lesion site with ShH10.CNTF/AAV2.shRNA-S1PR1 ( n = 6 mice) than with ShH10.CNTF/AAV2.GFP ( n = 5 mice) treatments (ANOVA, ∗ p

Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).

Techniques: CtB Assay, Infection, shRNA, Mouse Assay

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: Immunohistochemistry analysis of S1PR1 in hind limb muscle of sham and S aureus -infected mice. S1PR1 was significantly upregulated in the muscle of infected mice (red arrow) comparing with sham mice (green arrow), scale bar = 100 μ m.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Immunohistochemistry, Infection, Mouse Assay

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: MicroPET imaging of S1PR1 activity in S aureus -infected mice. (a) Radiosynthesis of S1PR1-specific radiotracer, [ 18 F]TZ4877; (b) representative sagittal microPET images of [ 18 F]TZ4877 in mice. Comparing with sham mice, the tracer uptake was significantly higher in the infected mice, and the increased uptake of the tracer showed S aureus dose dependent; (c) the tracer uptake in the brain was quantified; time-activity curves showed that the tracer uptake in infected mice was significantly higher than mice without infections; (d) the average tracer uptake in the brain from 30 to 50 min of the PET scan showed a dose-dependent manner. Data represent the mean ± SEM, n = 3 for each group.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Imaging, Activity Assay, Infection, Mouse Assay, Positron Emission Tomography

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: MicroPET imaging of S1PR1 activity in S aureus -infected mice. (a) Representative sagittal microPET images of [ 18 F]TZ4877 in the hind limb of mice. The tracer uptake was relatively low in the hind limb muscle with a SUV of ~1.5 in sham mice. Comparing with sham mice, the tracer uptake was significantly higher in the hind limb of infected mice; (b) time-activity curves showed that the tracer uptake in infected mice was significantly higher than sham mice; (c) the average tracer uptake in the hind limb muscle from 30 to 50 min of the PET scan showed a ~39% increase of SUV in infected mice with a P value of 0.0082. Data represent the mean ± SEM, n = 3 for each group.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Imaging, Activity Assay, Infection, Mouse Assay, Positron Emission Tomography

Journal: Oncotarget

Article Title: The novel sphingosine-1-phosphate receptors antagonist AD2900 affects lymphocyte activation and inhibits T-cell entry into the lymph nodes

doi: 10.18632/oncotarget.18626

Figure Lengend Snippet: A model for the intervention of AD2900 in the localization of murine T lymphocytes As an antagonist to S1P receptors 1–5, AD2900 can compete with S1P to bind S1P receptors leading to reduced S1P signaling and enhanced expression of S1P1 on T cells in S1P-rich environments such as the blood and the spleen. This altered expression, together with decreased CCR7 expression, inhibits T-cell entry into the lymph nodes (LNs) from the blood, causing accumulation of T cells in the blood. However, the entry of T cells to the spleen is not affected because it is not S1P dependent. Since Tcm-like cells express CCR7, these cells are attracted to the spleen and accumulate in it; yet, S1P1 elevated expression may have an effect on the S1P-dependent ingression of these cells from the MZ to the white pulp. Tef/em-like cells, which are CCR7 negative, are the primary T-cell subpopulation in the blood after AD2900 treatment. The significant decrease in naive T-cell counts in the circulation and peripheral lymphoid tissues tested may be explained by the inhibition of S1P signaling in the thymus leading to attenuated T-cell egression from the thymus to the circulation. Arrow key: thick = response; dashed = inhibition.

Article Snippet: To analyze S1P1 surface expression on AD2900-treated PBMCs, the following were used: anti-S1P1 (Alomone labs, Israel), anti-EDG-1 (Abcam, UK), Dylight405-conjugated AffiniPure Goat Anti-Rabbit lgG (H + L), and APC-conjugated AffiniPure F(ab')2 Goat Anti-Rabbit lgG(H + L) (Jackson ImmonoResearch, USA).

Techniques: Expressing, Inhibition

Journal: Oncotarget

Article Title: The novel sphingosine-1-phosphate receptors antagonist AD2900 affects lymphocyte activation and inhibits T-cell entry into the lymph nodes

doi: 10.18632/oncotarget.18626

Figure Lengend Snippet: The influence of AD2900 on S1P1- and CCR7-positive T-cell populations in blood, spleen, and peripheral lymph nodes C57BL/6 mice were orally administered with 1.8, 2.7, and 3.6 mg/l AD2900 or 1.8 mg/l FTY720 for 2 days, as shown in Figure 4 . Leukocytes from blood, spleen, and pLNs were collected and stained with CD3e and S1P1 or CCR7 fluorescent antibodies and then analyzed by FACS analysis. The percentages of S1P1+ CD3e+ T cells from blood (A) , spleen (B) , and pLNs (C) are shown. The percentages of CCR7+ CD3e+ T cells from blood (D) , spleen (E) , and pLNs (F) are shown. All the significances are compared to untreated healthy mice. Results summarize at least three independent experiments. Results of Student’s t -test:*(P

Article Snippet: To analyze S1P1 surface expression on AD2900-treated PBMCs, the following were used: anti-S1P1 (Alomone labs, Israel), anti-EDG-1 (Abcam, UK), Dylight405-conjugated AffiniPure Goat Anti-Rabbit lgG (H + L), and APC-conjugated AffiniPure F(ab')2 Goat Anti-Rabbit lgG(H + L) (Jackson ImmonoResearch, USA).

Techniques: Mouse Assay, Staining, FACS

Journal: Oncotarget

Article Title: The novel sphingosine-1-phosphate receptors antagonist AD2900 affects lymphocyte activation and inhibits T-cell entry into the lymph nodes

doi: 10.18632/oncotarget.18626

Figure Lengend Snippet: AD2900 downregulates the percentage of S1P1- and CCR7-expressing cells in PBMCs (A, B) The percentages of S1P1-positive PBMCs after the treatment with different concentrations of AD2900, FTY720, or SEW2871 and at different time points were examined by FACS analysis. S1P1 expression was tested in PBMCs after a 30-min treatment with AD2900 at different concentrations (A) or after a 30-min or 60-min treatment with 100 nM AD2900, FTY720, or SEW2871 (B) . (C) The percentage of CCR7-positive PBMCs was tested by FACS analysis after a 30-min treatment with 100 nM AD2900, FTY720, or SEW2871. All the significances are compared to untreated PBMCs. Results summarize the results of at least four independent experiments. Results of Student’s t -test: *(P

Article Snippet: To analyze S1P1 surface expression on AD2900-treated PBMCs, the following were used: anti-S1P1 (Alomone labs, Israel), anti-EDG-1 (Abcam, UK), Dylight405-conjugated AffiniPure Goat Anti-Rabbit lgG (H + L), and APC-conjugated AffiniPure F(ab')2 Goat Anti-Rabbit lgG(H + L) (Jackson ImmonoResearch, USA).

Techniques: Expressing, FACS