s1p assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    s1p assay kit - by Bioz Stars, 2023-01
    95/100 stars

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    1) Product Images from "Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells"

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102314

    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    Figure Legend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Techniques Used: Incubation, Purification, Enzyme-linked Immunosorbent Assay

    s1p levels  (Echelon Biosciences)


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    Echelon Biosciences s1p levels
    SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and <t>S1P</t> production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.
    S1p Levels, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1p levels - by Bioz Stars, 2023-01
    86/100 stars

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    1) Product Images from "Triptolide Inhibits the Biological Processes of HUVECs and HepG2 Cells via the Serine Palmitoyltransferase Long Chain Base Subunit 2/Sphingosine-1-Phosphate Signaling Pathway"

    Article Title: Triptolide Inhibits the Biological Processes of HUVECs and HepG2 Cells via the Serine Palmitoyltransferase Long Chain Base Subunit 2/Sphingosine-1-Phosphate Signaling Pathway

    Journal: Disease Markers

    doi: 10.1155/2022/9119423

    SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and S1P production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.
    Figure Legend Snippet: SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and S1P production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Techniques Used: Migration, Transfection, Small Interfering RNA, Negative Control, Plasmid Preparation, Expressing, Western Blot

    SPTLC2 can affect the proliferation, migration, invasion, and S1P production of HepG2 cells. After HepG2 cells were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), and SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), invasion (d), and S1P production (e) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.
    Figure Legend Snippet: SPTLC2 can affect the proliferation, migration, invasion, and S1P production of HepG2 cells. After HepG2 cells were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), and SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), invasion (d), and S1P production (e) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Techniques Used: Migration, Transfection, Small Interfering RNA, Negative Control, Plasmid Preparation, Expressing, Western Blot

    HUVECs may induce the proliferation, migration, and invasion of HepG2 cells via the S1P-S1PR pathway. (a) HUVECs and HepG2 cells were cocultured for 1 to 4 days, and a CCK-8 assay was used to determine HepG2 cells proliferation. HUVECs and HepG2 cells were cocultured for 24 h, and their migration (b) and invasion (c) were measured. HUVECs and HepG2 cells were cocultured for 1 to 4 days, and the S1P content (d) in the coculture system and the S1PR1, S1PR2, and S1PR3 protein expression (e) in HepG2 cells were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the noncoculture or control group.
    Figure Legend Snippet: HUVECs may induce the proliferation, migration, and invasion of HepG2 cells via the S1P-S1PR pathway. (a) HUVECs and HepG2 cells were cocultured for 1 to 4 days, and a CCK-8 assay was used to determine HepG2 cells proliferation. HUVECs and HepG2 cells were cocultured for 24 h, and their migration (b) and invasion (c) were measured. HUVECs and HepG2 cells were cocultured for 1 to 4 days, and the S1P content (d) in the coculture system and the S1PR1, S1PR2, and S1PR3 protein expression (e) in HepG2 cells were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the noncoculture or control group.

    Techniques Used: Migration, CCK-8 Assay, Expressing

    sphingosine 1 phoshate s1p  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phoshate s1p
    <t> Sphingosine-1-phosphate </t> levels in HDL, HDL2, and HDL3.
    Sphingosine 1 Phoshate S1p, supplied by Echelon Biosciences, 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/sphingosine 1 phoshate s1p/product/Echelon Biosciences
    Average 86 stars, based on 1 article reviews
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    sphingosine 1 phoshate s1p - by Bioz Stars, 2023-01
    86/100 stars

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    1) Product Images from "eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency"

    Article Title: eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095925

     Sphingosine-1-phosphate  levels in HDL, HDL2, and HDL3.
    Figure Legend Snippet: Sphingosine-1-phosphate levels in HDL, HDL2, and HDL3.

    Techniques Used:

    anti s1p  (Echelon Biosciences)


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    Echelon Biosciences anti s1p
    Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with <t>S1P.</t> Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.
    Anti S1p, supplied by Echelon Biosciences, 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/anti s1p/product/Echelon Biosciences
    Average 86 stars, based on 1 article reviews
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    anti s1p - by Bioz Stars, 2023-01
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    1) Product Images from "Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients"

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms232416117

    Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with S1P. Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.
    Figure Legend Snippet: Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with S1P. Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Techniques Used: Activation Assay, Flow Cytometry, Expressing, Positive Control

    Calcium flux assay. Changes in basophil cytosolic Ca 2+ levels after stimulation with S1P. Purified basophils were incubated with Fluo-4 AM (4 µM), and calcium flux was measured by flow cytometry. ( a ) Differences in relative cytosolic Ca 2+ concentrations to baseline, after stimulation with 500 nM ionomycin (positive control), 0.1, 1, or 10 µM S1P ( n = 6). ( b ) Changes in relative cytosolic Ca 2+ concentrations after priming with IL-3 (10 ng/mL) and stimulation with either ionomycin or different concentrations of S1P ( n = 4). ( c ) Changes in fluorescence over 300 s during application of RPMI, ionomycin, and 10 µM S1P. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.
    Figure Legend Snippet: Calcium flux assay. Changes in basophil cytosolic Ca 2+ levels after stimulation with S1P. Purified basophils were incubated with Fluo-4 AM (4 µM), and calcium flux was measured by flow cytometry. ( a ) Differences in relative cytosolic Ca 2+ concentrations to baseline, after stimulation with 500 nM ionomycin (positive control), 0.1, 1, or 10 µM S1P ( n = 6). ( b ) Changes in relative cytosolic Ca 2+ concentrations after priming with IL-3 (10 ng/mL) and stimulation with either ionomycin or different concentrations of S1P ( n = 4). ( c ) Changes in fluorescence over 300 s during application of RPMI, ionomycin, and 10 µM S1P. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Techniques Used: Calcium Flux Assay, Purification, Incubation, Flow Cytometry, Positive Control, Fluorescence

    Apoptosis assay. Percentage of viable and combined apoptotic and late apoptotic basophils after S1P stimulation. Flow cytometry analysis of the annexin V-FITC and PI apoptosis assay. Basophils were stimulated with fatty acid free BSA (Co), S1P (0.1 µM, 1 µM, 10 µM), or staurosporine (1 µM) for 24 h ( n = 3). ( a ) Percentage of viable basophils. ( b ) Percentage of combined apoptotic and late apoptotic basophils. Stimulation with 10 µM S1P decreases the number of viable and increases the number of apoptotic cells. 0.1 µM S1P reduces the number of apoptotic basophils. ( c ) Representative dot plots demonstrate the different stages of apoptosis for Co, Ionomycin, and 10 µM S1P stimulated basophils. Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co group.
    Figure Legend Snippet: Apoptosis assay. Percentage of viable and combined apoptotic and late apoptotic basophils after S1P stimulation. Flow cytometry analysis of the annexin V-FITC and PI apoptosis assay. Basophils were stimulated with fatty acid free BSA (Co), S1P (0.1 µM, 1 µM, 10 µM), or staurosporine (1 µM) for 24 h ( n = 3). ( a ) Percentage of viable basophils. ( b ) Percentage of combined apoptotic and late apoptotic basophils. Stimulation with 10 µM S1P decreases the number of viable and increases the number of apoptotic cells. 0.1 µM S1P reduces the number of apoptotic basophils. ( c ) Representative dot plots demonstrate the different stages of apoptosis for Co, Ionomycin, and 10 µM S1P stimulated basophils. Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co group.

    Techniques Used: Apoptosis Assay, Flow Cytometry

    Chemotaxis assay and S1PR1 protein expression. Basophil chemotactic activity and flow cytometry analysis of S1PR1 protein expression in NA and AT patients. NA = non-atopic, AT = atopic, SSC = side scatter. Basophils were placed in the upper chamber of a transwell migration assay, and chemotactic activity toward stimuli in the bottom chamber was measured. Cells were counted after 3 h through flow cytometry. Fatty acid free BSA (Co), S1P (1 and 10 µM), or eotaxin (Eot) (8 ng/mL) were applied in FCS-free RPMI. The chemotactic index was calculated in relation to the transmigrated cells in the Co group. ( a ) Chemotactic index of NA basophils ( n = 6). Basophils are significantly attracted to eotaxin. ( b ) Chemotactic index of AT basophils ( n = 3). Basophils migrate towards eotaxin, whereas 1 µM S1P significantly reduces the number of transmigrating cells. ( c ) Flow cytometry analysis of protein level S1PR1 expression. Percent of S1PR1 positive basophils of NA ( n = 6) and AT ( n = 5) patients. AT patients express significantly less S1PR1 on the cell surface than the NA group. ( d ) Flow cytometry analysis of S1PR1 MFI in NA ( n = 6) and AT ( n = 5) basophils. ( e ) Representative dot plot of S1PR1 protein expression in NA basophils (top) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained NA basophils (bottom). ( f ) Representative dot plot of S1PR1 protein expression in AT basophils ( top ) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained AT basophils ( bottom ). Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co or NA group.
    Figure Legend Snippet: Chemotaxis assay and S1PR1 protein expression. Basophil chemotactic activity and flow cytometry analysis of S1PR1 protein expression in NA and AT patients. NA = non-atopic, AT = atopic, SSC = side scatter. Basophils were placed in the upper chamber of a transwell migration assay, and chemotactic activity toward stimuli in the bottom chamber was measured. Cells were counted after 3 h through flow cytometry. Fatty acid free BSA (Co), S1P (1 and 10 µM), or eotaxin (Eot) (8 ng/mL) were applied in FCS-free RPMI. The chemotactic index was calculated in relation to the transmigrated cells in the Co group. ( a ) Chemotactic index of NA basophils ( n = 6). Basophils are significantly attracted to eotaxin. ( b ) Chemotactic index of AT basophils ( n = 3). Basophils migrate towards eotaxin, whereas 1 µM S1P significantly reduces the number of transmigrating cells. ( c ) Flow cytometry analysis of protein level S1PR1 expression. Percent of S1PR1 positive basophils of NA ( n = 6) and AT ( n = 5) patients. AT patients express significantly less S1PR1 on the cell surface than the NA group. ( d ) Flow cytometry analysis of S1PR1 MFI in NA ( n = 6) and AT ( n = 5) basophils. ( e ) Representative dot plot of S1PR1 protein expression in NA basophils (top) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained NA basophils (bottom). ( f ) Representative dot plot of S1PR1 protein expression in AT basophils ( top ) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained AT basophils ( bottom ). Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co or NA group.

    Techniques Used: Chemotaxis Assay, Expressing, Activity Assay, Flow Cytometry, Transwell Migration Assay, Staining

    S1P immunofluorescence staining of S1P positive basophils in AD skin. AD = atopic dermatitis. A lesional AD skin biopsy was fixed in 4% PFA, and a double immunofluorescence staining was performed. Antibodies against the basophil marker 2D7 (red) and the lipid S1P (green) were utilized. Nuclei were stained with DAPI (blue). The image was acquired through fluorescence microscopy at 40× magnification. The dotted line marks the junction of the epidermis (top) and dermis (bottom), and arrows point to intradermal basophils. Basophils in the dermis possess intracellular S1P.
    Figure Legend Snippet: S1P immunofluorescence staining of S1P positive basophils in AD skin. AD = atopic dermatitis. A lesional AD skin biopsy was fixed in 4% PFA, and a double immunofluorescence staining was performed. Antibodies against the basophil marker 2D7 (red) and the lipid S1P (green) were utilized. Nuclei were stained with DAPI (blue). The image was acquired through fluorescence microscopy at 40× magnification. The dotted line marks the junction of the epidermis (top) and dermis (bottom), and arrows point to intradermal basophils. Basophils in the dermis possess intracellular S1P.

    Techniques Used: Immunofluorescence, Staining, Double Immunofluorescence Staining, Marker, Fluorescence, Microscopy

    s1p  (Echelon Biosciences)


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    Echelon Biosciences s1p
    SPHK1 expression was upregulated in TNBC patients, and <t>S1P</t> levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.
    S1p, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    s1p - by Bioz Stars, 2023-01
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    1) Product Images from "Triple Negative Breast Cancer Depends on Sphingosine Kinase 1 (SphK1)/Sphingosine-1-Phosphate (S1P)/Sphingosine 1-Phosphate Receptor 3 (S1PR3)/Notch Signaling for Metastasis"

    Article Title: Triple Negative Breast Cancer Depends on Sphingosine Kinase 1 (SphK1)/Sphingosine-1-Phosphate (S1P)/Sphingosine 1-Phosphate Receptor 3 (S1PR3)/Notch Signaling for Metastasis

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.905833

    SPHK1 expression was upregulated in TNBC patients, and S1P levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.
    Figure Legend Snippet: SPHK1 expression was upregulated in TNBC patients, and S1P levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.

    Techniques Used: Expressing, Immunohistochemistry

    SphK1 leads to increase S1P levels and enhances Notch signaling pathway via S1PR3. ( A ) TNBC cells MDA-MB-231 were transfected with Ad-NC-siRNA or Ad-SPHK1-siRNA #2 at 10 MOI for 24 hours, RT-PCR assay was using for detection the expression of Hes1 (Notch signaling target gene), Gli1 (Hedgehog signaling target gene), and Dkk1 (Wnt signaling target gene). ( B ) MDA-MB-231 cells were treatment with S1P (200 nM) or LPA (200 nM) for 24 hours, expression levels of Hes1, Gli1, and Dkk1 were detected by RT-PCR. ( C ) Effects of TY52156 (2 mM), CAY10444 (5 mM) or JTE013 (2 mM) on S1P-induced Hes1 expression by RT-PCR. Data represent mean ±SD (n=3). ( D ) Effects of S1P, LPA, and N1ICD overexpression (N1ICD OE) on N1ICD production by western blotting, GAPDH was regard as control.
    Figure Legend Snippet: SphK1 leads to increase S1P levels and enhances Notch signaling pathway via S1PR3. ( A ) TNBC cells MDA-MB-231 were transfected with Ad-NC-siRNA or Ad-SPHK1-siRNA #2 at 10 MOI for 24 hours, RT-PCR assay was using for detection the expression of Hes1 (Notch signaling target gene), Gli1 (Hedgehog signaling target gene), and Dkk1 (Wnt signaling target gene). ( B ) MDA-MB-231 cells were treatment with S1P (200 nM) or LPA (200 nM) for 24 hours, expression levels of Hes1, Gli1, and Dkk1 were detected by RT-PCR. ( C ) Effects of TY52156 (2 mM), CAY10444 (5 mM) or JTE013 (2 mM) on S1P-induced Hes1 expression by RT-PCR. Data represent mean ±SD (n=3). ( D ) Effects of S1P, LPA, and N1ICD overexpression (N1ICD OE) on N1ICD production by western blotting, GAPDH was regard as control.

    Techniques Used: Transfection, Reverse Transcription Polymerase Chain Reaction, Expressing, Over Expression, Western Blot

    s1p competitive elisa kit  (Echelon Biosciences)


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    Echelon Biosciences s1p competitive elisa kit
    S1p Competitive Elisa Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    s1p competitive elisa kit - by Bioz Stars, 2023-01
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    s1p assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/s1p assay kit/product/Echelon Biosciences
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1p assay kit - by Bioz Stars, 2023-01
    95/100 stars

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    1) Product Images from "Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells"

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102314

    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    Figure Legend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Techniques Used: Incubation, Purification, Enzyme-linked Immunosorbent Assay

    s1p  (Echelon Biosciences)


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    Echelon Biosciences s1p
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells"

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102314

    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    Figure Legend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Techniques Used: Incubation, Purification, Enzyme-linked Immunosorbent Assay

    s1p  (Echelon Biosciences)


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    Echelon Biosciences s1p
    Elevating <t>S1P</t> levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, <t>sphingosine-1-phoshate;</t> SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    S1p, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice"

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    Journal: Skeletal Muscle

    doi: 10.1186/2044-5040-3-20

    Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    Figure Legend Snippet: Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Techniques Used: Staining

    S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.
    Figure Legend Snippet: S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Techniques Used: Functional Assay, Injection, In Vitro, Isolation, Incubation, Concentration Assay

    Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: Injection, Staining

    Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: In Vivo, Injection, Western Blot

    Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: Injection, Activation Assay, Liquid Chromatography with Mass Spectroscopy, Staining, Liquid Chromatography, Mass Spectrometry

    IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.
    Figure Legend Snippet: IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Techniques Used: Injection, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry

    Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    Figure Legend Snippet: Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Techniques Used: Activation Assay

    s1p biotin  (Echelon Biosciences)


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    Echelon Biosciences s1p biotin
    Elevating <t>S1P</t> levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, <t>sphingosine-1-phoshate;</t> SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    S1p Biotin, supplied by Echelon Biosciences, 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 biotin/product/Echelon Biosciences
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1p biotin - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice"

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    Journal: Skeletal Muscle

    doi: 10.1186/2044-5040-3-20

    Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    Figure Legend Snippet: Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Techniques Used: Staining

    S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.
    Figure Legend Snippet: S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Techniques Used: Functional Assay, Injection, In Vitro, Isolation, Incubation, Concentration Assay

    Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: Injection, Staining

    Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: In Vivo, Injection, Western Blot

    Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.
    Figure Legend Snippet: Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.

    Techniques Used: Injection, Activation Assay, Liquid Chromatography with Mass Spectroscopy, Staining, Liquid Chromatography, Mass Spectrometry

    IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.
    Figure Legend Snippet: IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Techniques Used: Injection, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry

    Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
    Figure Legend Snippet: Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Techniques Used: Activation Assay

    s1p  (Echelon Biosciences)


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    Structured Review

    Echelon Biosciences s1p
    <t>S1P</t> concentration in BALF collected on the seventh day after intratracheal administration of bleomycin or non-treated control. There were no significant differences in S1P concentration in BALF between wild-type (WT) and knockout (KO) mice at baseline or on the seventh day after bleomycin administration, nor were there significant differences in the S1P concentrations in BALF collected before and after bleomycin administration in WT (baseline vs. seventh day; p = 0.14) or KO (baseline vs. seventh day; p = 0.50) mice.
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    Images

    1) Product Images from "Knock Out of S1P3 Receptor Signaling Attenuates Inflammation and Fibrosis in Bleomycin-Induced Lung Injury Mice Model"

    Article Title: Knock Out of S1P3 Receptor Signaling Attenuates Inflammation and Fibrosis in Bleomycin-Induced Lung Injury Mice Model

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0106792

    S1P concentration in BALF collected on the seventh day after intratracheal administration of bleomycin or non-treated control. There were no significant differences in S1P concentration in BALF between wild-type (WT) and knockout (KO) mice at baseline or on the seventh day after bleomycin administration, nor were there significant differences in the S1P concentrations in BALF collected before and after bleomycin administration in WT (baseline vs. seventh day; p = 0.14) or KO (baseline vs. seventh day; p = 0.50) mice.
    Figure Legend Snippet: S1P concentration in BALF collected on the seventh day after intratracheal administration of bleomycin or non-treated control. There were no significant differences in S1P concentration in BALF between wild-type (WT) and knockout (KO) mice at baseline or on the seventh day after bleomycin administration, nor were there significant differences in the S1P concentrations in BALF collected before and after bleomycin administration in WT (baseline vs. seventh day; p = 0.14) or KO (baseline vs. seventh day; p = 0.50) mice.

    Techniques Used: Concentration Assay, Knock-Out

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    Echelon Biosciences s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Echelon Biosciences s1p levels
    SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and <t>S1P</t> production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.
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    Echelon Biosciences sphingosine 1 phoshate s1p
    <t> Sphingosine-1-phosphate </t> levels in HDL, HDL2, and HDL3.
    Sphingosine 1 Phoshate S1p, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Echelon Biosciences anti s1p
    Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with <t>S1P.</t> Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.
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    Echelon Biosciences s1p
    SPHK1 expression was upregulated in TNBC patients, and <t>S1P</t> levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.
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    Echelon Biosciences s1p competitive elisa kit
    SPHK1 expression was upregulated in TNBC patients, and <t>S1P</t> levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.
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    Echelon Biosciences s1p biotin
    Elevating <t>S1P</t> levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, <t>sphingosine-1-phoshate;</t> SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.
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    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Journal: PLoS ONE

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    doi: 10.1371/journal.pone.0102314

    Figure Lengend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Article Snippet: Concentrations of S1P in culture supernatants and pDMVEC cell lysates were determined using the S1P Assay Kit (Echelon) according to the manufacturers’ instructions.

    Techniques: Incubation, Purification, Enzyme-linked Immunosorbent Assay

    SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and S1P production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Journal: Disease Markers

    Article Title: Triptolide Inhibits the Biological Processes of HUVECs and HepG2 Cells via the Serine Palmitoyltransferase Long Chain Base Subunit 2/Sphingosine-1-Phosphate Signaling Pathway

    doi: 10.1155/2022/9119423

    Figure Lengend Snippet: SPTLC2 can affect the proliferation, migration, adhesion, angiogenesis, and S1P production of HUVECs. After HUVECs were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), or SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), adhesion (d), angiogenesis (e), and S1P production (f) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Article Snippet: S1P levels were detected by a sphingosine-1-phosphate ELISA kit (Echelon, Salt Lake, USA) according to the manufacturer's protocol.

    Techniques: Migration, Transfection, Small Interfering RNA, Negative Control, Plasmid Preparation, Expressing, Western Blot

    SPTLC2 can affect the proliferation, migration, invasion, and S1P production of HepG2 cells. After HepG2 cells were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), and SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), invasion (d), and S1P production (e) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Journal: Disease Markers

    Article Title: Triptolide Inhibits the Biological Processes of HUVECs and HepG2 Cells via the Serine Palmitoyltransferase Long Chain Base Subunit 2/Sphingosine-1-Phosphate Signaling Pathway

    doi: 10.1155/2022/9119423

    Figure Lengend Snippet: SPTLC2 can affect the proliferation, migration, invasion, and S1P production of HepG2 cells. After HepG2 cells were transfected with siR-NC (SPTLC2 small interfering RNA negative control), siR-SPTLC2 (SPTLC2 small interfering RNA), SPTLC2-NC (SPTLC2 plasmid negative control), and SPTLC2 (SPTLC2 plasmid), the protein expression of SPTLC2 was detected by western blotting assay (a), and the proliferation (b), migration (c), invasion (d), and S1P production (e) were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the blank group.

    Article Snippet: S1P levels were detected by a sphingosine-1-phosphate ELISA kit (Echelon, Salt Lake, USA) according to the manufacturer's protocol.

    Techniques: Migration, Transfection, Small Interfering RNA, Negative Control, Plasmid Preparation, Expressing, Western Blot

    HUVECs may induce the proliferation, migration, and invasion of HepG2 cells via the S1P-S1PR pathway. (a) HUVECs and HepG2 cells were cocultured for 1 to 4 days, and a CCK-8 assay was used to determine HepG2 cells proliferation. HUVECs and HepG2 cells were cocultured for 24 h, and their migration (b) and invasion (c) were measured. HUVECs and HepG2 cells were cocultured for 1 to 4 days, and the S1P content (d) in the coculture system and the S1PR1, S1PR2, and S1PR3 protein expression (e) in HepG2 cells were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the noncoculture or control group.

    Journal: Disease Markers

    Article Title: Triptolide Inhibits the Biological Processes of HUVECs and HepG2 Cells via the Serine Palmitoyltransferase Long Chain Base Subunit 2/Sphingosine-1-Phosphate Signaling Pathway

    doi: 10.1155/2022/9119423

    Figure Lengend Snippet: HUVECs may induce the proliferation, migration, and invasion of HepG2 cells via the S1P-S1PR pathway. (a) HUVECs and HepG2 cells were cocultured for 1 to 4 days, and a CCK-8 assay was used to determine HepG2 cells proliferation. HUVECs and HepG2 cells were cocultured for 24 h, and their migration (b) and invasion (c) were measured. HUVECs and HepG2 cells were cocultured for 1 to 4 days, and the S1P content (d) in the coculture system and the S1PR1, S1PR2, and S1PR3 protein expression (e) in HepG2 cells were measured. ∗ P < 0.05, ∗∗ P < 0.01 versus the noncoculture or control group.

    Article Snippet: S1P levels were detected by a sphingosine-1-phosphate ELISA kit (Echelon, Salt Lake, USA) according to the manufacturer's protocol.

    Techniques: Migration, CCK-8 Assay, Expressing

     Sphingosine-1-phosphate  levels in HDL, HDL2, and HDL3.

    Journal: PLoS ONE

    Article Title: eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency

    doi: 10.1371/journal.pone.0095925

    Figure Lengend Snippet: Sphingosine-1-phosphate levels in HDL, HDL2, and HDL3.

    Article Snippet: The concentration of sphingosine-1-phoshate (S1P) in isolated HDL fractions was measured with a commercial competitive ELISA kit (Echelon Biosciences Inc., Salt Lake City, UT, USA) and normalized by protein concentration.

    Techniques:

    Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with S1P. Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Journal: International Journal of Molecular Sciences

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    doi: 10.3390/ijms232416117

    Figure Lengend Snippet: Basophil activation test assay. Flow cytometry analysis of CD63 & CD203c protein expressions after stimulation with S1P. Basophils were stimulated with fatty acid free BSA (4 mg/mL) (Co), S1P (0.1 µM, 1 µM, 10 µM) or a-IgE (1 µg/mL) for 30 min ( n = 5). ( a ) CD63 protein expression is significantly increased for the positive control a-IgE. ( b ) CD203c expression is markedly higher in a-IgE than in Co-stimulated cells. ( c ) Representative overlay histogram of CD63 expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. ( d ) Representative overlay histogram of CD203c expression in Co (dark grey) and a-IgE stimulated (light grey) basophils. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Article Snippet: Slides were consecutively incubated anti-S1P (1:200) (Z-P300) (Echelon Biosciences, Salt Lake City, UT, USA), goat anti-mouse Alexa Fluor 488 secondary antibody (1:2000) (15607878) (ThermoFisher Scientific, Waltham, MA, USA) and the basophil specific anti-2D7 Alexa Fluor 647 (RRID: AB_1967142) (1:100) (BioLegend, San Diego, CA, USA).

    Techniques: Activation Assay, Flow Cytometry, Expressing, Positive Control

    Calcium flux assay. Changes in basophil cytosolic Ca 2+ levels after stimulation with S1P. Purified basophils were incubated with Fluo-4 AM (4 µM), and calcium flux was measured by flow cytometry. ( a ) Differences in relative cytosolic Ca 2+ concentrations to baseline, after stimulation with 500 nM ionomycin (positive control), 0.1, 1, or 10 µM S1P ( n = 6). ( b ) Changes in relative cytosolic Ca 2+ concentrations after priming with IL-3 (10 ng/mL) and stimulation with either ionomycin or different concentrations of S1P ( n = 4). ( c ) Changes in fluorescence over 300 s during application of RPMI, ionomycin, and 10 µM S1P. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Journal: International Journal of Molecular Sciences

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    doi: 10.3390/ijms232416117

    Figure Lengend Snippet: Calcium flux assay. Changes in basophil cytosolic Ca 2+ levels after stimulation with S1P. Purified basophils were incubated with Fluo-4 AM (4 µM), and calcium flux was measured by flow cytometry. ( a ) Differences in relative cytosolic Ca 2+ concentrations to baseline, after stimulation with 500 nM ionomycin (positive control), 0.1, 1, or 10 µM S1P ( n = 6). ( b ) Changes in relative cytosolic Ca 2+ concentrations after priming with IL-3 (10 ng/mL) and stimulation with either ionomycin or different concentrations of S1P ( n = 4). ( c ) Changes in fluorescence over 300 s during application of RPMI, ionomycin, and 10 µM S1P. Data are shown as mean + SEM, p -values (*** ≤ 0.001). Significances are calculated in relation to the Co group.

    Article Snippet: Slides were consecutively incubated anti-S1P (1:200) (Z-P300) (Echelon Biosciences, Salt Lake City, UT, USA), goat anti-mouse Alexa Fluor 488 secondary antibody (1:2000) (15607878) (ThermoFisher Scientific, Waltham, MA, USA) and the basophil specific anti-2D7 Alexa Fluor 647 (RRID: AB_1967142) (1:100) (BioLegend, San Diego, CA, USA).

    Techniques: Calcium Flux Assay, Purification, Incubation, Flow Cytometry, Positive Control, Fluorescence

    Apoptosis assay. Percentage of viable and combined apoptotic and late apoptotic basophils after S1P stimulation. Flow cytometry analysis of the annexin V-FITC and PI apoptosis assay. Basophils were stimulated with fatty acid free BSA (Co), S1P (0.1 µM, 1 µM, 10 µM), or staurosporine (1 µM) for 24 h ( n = 3). ( a ) Percentage of viable basophils. ( b ) Percentage of combined apoptotic and late apoptotic basophils. Stimulation with 10 µM S1P decreases the number of viable and increases the number of apoptotic cells. 0.1 µM S1P reduces the number of apoptotic basophils. ( c ) Representative dot plots demonstrate the different stages of apoptosis for Co, Ionomycin, and 10 µM S1P stimulated basophils. Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co group.

    Journal: International Journal of Molecular Sciences

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    doi: 10.3390/ijms232416117

    Figure Lengend Snippet: Apoptosis assay. Percentage of viable and combined apoptotic and late apoptotic basophils after S1P stimulation. Flow cytometry analysis of the annexin V-FITC and PI apoptosis assay. Basophils were stimulated with fatty acid free BSA (Co), S1P (0.1 µM, 1 µM, 10 µM), or staurosporine (1 µM) for 24 h ( n = 3). ( a ) Percentage of viable basophils. ( b ) Percentage of combined apoptotic and late apoptotic basophils. Stimulation with 10 µM S1P decreases the number of viable and increases the number of apoptotic cells. 0.1 µM S1P reduces the number of apoptotic basophils. ( c ) Representative dot plots demonstrate the different stages of apoptosis for Co, Ionomycin, and 10 µM S1P stimulated basophils. Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co group.

    Article Snippet: Slides were consecutively incubated anti-S1P (1:200) (Z-P300) (Echelon Biosciences, Salt Lake City, UT, USA), goat anti-mouse Alexa Fluor 488 secondary antibody (1:2000) (15607878) (ThermoFisher Scientific, Waltham, MA, USA) and the basophil specific anti-2D7 Alexa Fluor 647 (RRID: AB_1967142) (1:100) (BioLegend, San Diego, CA, USA).

    Techniques: Apoptosis Assay, Flow Cytometry

    Chemotaxis assay and S1PR1 protein expression. Basophil chemotactic activity and flow cytometry analysis of S1PR1 protein expression in NA and AT patients. NA = non-atopic, AT = atopic, SSC = side scatter. Basophils were placed in the upper chamber of a transwell migration assay, and chemotactic activity toward stimuli in the bottom chamber was measured. Cells were counted after 3 h through flow cytometry. Fatty acid free BSA (Co), S1P (1 and 10 µM), or eotaxin (Eot) (8 ng/mL) were applied in FCS-free RPMI. The chemotactic index was calculated in relation to the transmigrated cells in the Co group. ( a ) Chemotactic index of NA basophils ( n = 6). Basophils are significantly attracted to eotaxin. ( b ) Chemotactic index of AT basophils ( n = 3). Basophils migrate towards eotaxin, whereas 1 µM S1P significantly reduces the number of transmigrating cells. ( c ) Flow cytometry analysis of protein level S1PR1 expression. Percent of S1PR1 positive basophils of NA ( n = 6) and AT ( n = 5) patients. AT patients express significantly less S1PR1 on the cell surface than the NA group. ( d ) Flow cytometry analysis of S1PR1 MFI in NA ( n = 6) and AT ( n = 5) basophils. ( e ) Representative dot plot of S1PR1 protein expression in NA basophils (top) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained NA basophils (bottom). ( f ) Representative dot plot of S1PR1 protein expression in AT basophils ( top ) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained AT basophils ( bottom ). Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co or NA group.

    Journal: International Journal of Molecular Sciences

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    doi: 10.3390/ijms232416117

    Figure Lengend Snippet: Chemotaxis assay and S1PR1 protein expression. Basophil chemotactic activity and flow cytometry analysis of S1PR1 protein expression in NA and AT patients. NA = non-atopic, AT = atopic, SSC = side scatter. Basophils were placed in the upper chamber of a transwell migration assay, and chemotactic activity toward stimuli in the bottom chamber was measured. Cells were counted after 3 h through flow cytometry. Fatty acid free BSA (Co), S1P (1 and 10 µM), or eotaxin (Eot) (8 ng/mL) were applied in FCS-free RPMI. The chemotactic index was calculated in relation to the transmigrated cells in the Co group. ( a ) Chemotactic index of NA basophils ( n = 6). Basophils are significantly attracted to eotaxin. ( b ) Chemotactic index of AT basophils ( n = 3). Basophils migrate towards eotaxin, whereas 1 µM S1P significantly reduces the number of transmigrating cells. ( c ) Flow cytometry analysis of protein level S1PR1 expression. Percent of S1PR1 positive basophils of NA ( n = 6) and AT ( n = 5) patients. AT patients express significantly less S1PR1 on the cell surface than the NA group. ( d ) Flow cytometry analysis of S1PR1 MFI in NA ( n = 6) and AT ( n = 5) basophils. ( e ) Representative dot plot of S1PR1 protein expression in NA basophils (top) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained NA basophils (bottom). ( f ) Representative dot plot of S1PR1 protein expression in AT basophils ( top ) and representative histogram of S1PR1 (light grey) and isotype control (dark grey) stained AT basophils ( bottom ). Data shown as mean + SEM, p -values (* ≤ 0.05; ** ≤ 0.01). Significances are calculated in relation to the Co or NA group.

    Article Snippet: Slides were consecutively incubated anti-S1P (1:200) (Z-P300) (Echelon Biosciences, Salt Lake City, UT, USA), goat anti-mouse Alexa Fluor 488 secondary antibody (1:2000) (15607878) (ThermoFisher Scientific, Waltham, MA, USA) and the basophil specific anti-2D7 Alexa Fluor 647 (RRID: AB_1967142) (1:100) (BioLegend, San Diego, CA, USA).

    Techniques: Chemotaxis Assay, Expressing, Activity Assay, Flow Cytometry, Transwell Migration Assay, Staining

    S1P immunofluorescence staining of S1P positive basophils in AD skin. AD = atopic dermatitis. A lesional AD skin biopsy was fixed in 4% PFA, and a double immunofluorescence staining was performed. Antibodies against the basophil marker 2D7 (red) and the lipid S1P (green) were utilized. Nuclei were stained with DAPI (blue). The image was acquired through fluorescence microscopy at 40× magnification. The dotted line marks the junction of the epidermis (top) and dermis (bottom), and arrows point to intradermal basophils. Basophils in the dermis possess intracellular S1P.

    Journal: International Journal of Molecular Sciences

    Article Title: Differential Upregulation and Functional Activity of S1PR1 in Human Peripheral Blood Basophils of Atopic Patients

    doi: 10.3390/ijms232416117

    Figure Lengend Snippet: S1P immunofluorescence staining of S1P positive basophils in AD skin. AD = atopic dermatitis. A lesional AD skin biopsy was fixed in 4% PFA, and a double immunofluorescence staining was performed. Antibodies against the basophil marker 2D7 (red) and the lipid S1P (green) were utilized. Nuclei were stained with DAPI (blue). The image was acquired through fluorescence microscopy at 40× magnification. The dotted line marks the junction of the epidermis (top) and dermis (bottom), and arrows point to intradermal basophils. Basophils in the dermis possess intracellular S1P.

    Article Snippet: Slides were consecutively incubated anti-S1P (1:200) (Z-P300) (Echelon Biosciences, Salt Lake City, UT, USA), goat anti-mouse Alexa Fluor 488 secondary antibody (1:2000) (15607878) (ThermoFisher Scientific, Waltham, MA, USA) and the basophil specific anti-2D7 Alexa Fluor 647 (RRID: AB_1967142) (1:100) (BioLegend, San Diego, CA, USA).

    Techniques: Immunofluorescence, Staining, Double Immunofluorescence Staining, Marker, Fluorescence, Microscopy

    SPHK1 expression was upregulated in TNBC patients, and S1P levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Triple Negative Breast Cancer Depends on Sphingosine Kinase 1 (SphK1)/Sphingosine-1-Phosphate (S1P)/Sphingosine 1-Phosphate Receptor 3 (S1PR3)/Notch Signaling for Metastasis

    doi: 10.12659/MSM.905833

    Figure Lengend Snippet: SPHK1 expression was upregulated in TNBC patients, and S1P levels are correlated with high expression of pSphK1. ( A ) The mRNA expression of SPHK1 in HER2-positive, HER2 (+), HER2-negative, HER2 (−), PR-positive, PR (+), PR-negative, PR (−) and ER-positive, ER (+), ER-negative, ER (−) breast cancer patients. ( B ) The positive ratio of pSphK1 was detected in HER2-positive and negative patients. ( C ) The expression of HER2 and pSphK1 were detected by immunohistochemistry, the represent images were shown. ( D ) S1P levels were detected in breast cancer patients with pSphK1 positive or negative.

    Article Snippet: The levels of S1P were detected by S1P competitive ELISA kit, which had a sensitivity of 30 nM (Echelon Bioscience Inc.).

    Techniques: Expressing, Immunohistochemistry

    SphK1 leads to increase S1P levels and enhances Notch signaling pathway via S1PR3. ( A ) TNBC cells MDA-MB-231 were transfected with Ad-NC-siRNA or Ad-SPHK1-siRNA #2 at 10 MOI for 24 hours, RT-PCR assay was using for detection the expression of Hes1 (Notch signaling target gene), Gli1 (Hedgehog signaling target gene), and Dkk1 (Wnt signaling target gene). ( B ) MDA-MB-231 cells were treatment with S1P (200 nM) or LPA (200 nM) for 24 hours, expression levels of Hes1, Gli1, and Dkk1 were detected by RT-PCR. ( C ) Effects of TY52156 (2 mM), CAY10444 (5 mM) or JTE013 (2 mM) on S1P-induced Hes1 expression by RT-PCR. Data represent mean ±SD (n=3). ( D ) Effects of S1P, LPA, and N1ICD overexpression (N1ICD OE) on N1ICD production by western blotting, GAPDH was regard as control.

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Triple Negative Breast Cancer Depends on Sphingosine Kinase 1 (SphK1)/Sphingosine-1-Phosphate (S1P)/Sphingosine 1-Phosphate Receptor 3 (S1PR3)/Notch Signaling for Metastasis

    doi: 10.12659/MSM.905833

    Figure Lengend Snippet: SphK1 leads to increase S1P levels and enhances Notch signaling pathway via S1PR3. ( A ) TNBC cells MDA-MB-231 were transfected with Ad-NC-siRNA or Ad-SPHK1-siRNA #2 at 10 MOI for 24 hours, RT-PCR assay was using for detection the expression of Hes1 (Notch signaling target gene), Gli1 (Hedgehog signaling target gene), and Dkk1 (Wnt signaling target gene). ( B ) MDA-MB-231 cells were treatment with S1P (200 nM) or LPA (200 nM) for 24 hours, expression levels of Hes1, Gli1, and Dkk1 were detected by RT-PCR. ( C ) Effects of TY52156 (2 mM), CAY10444 (5 mM) or JTE013 (2 mM) on S1P-induced Hes1 expression by RT-PCR. Data represent mean ±SD (n=3). ( D ) Effects of S1P, LPA, and N1ICD overexpression (N1ICD OE) on N1ICD production by western blotting, GAPDH was regard as control.

    Article Snippet: The levels of S1P were detected by S1P competitive ELISA kit, which had a sensitivity of 30 nM (Echelon Bioscience Inc.).

    Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Expressing, Over Expression, Western Blot

    Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: Elevating S1P levels with THI increases muscle fiber size. ( A ) Staining for laminin (green) and DAPI (blue) depict a dramatic increase in muscle fiber size in both injured and uninjured quadriceps (quads) with THI treatment. Depicted are quadriceps muscles from 11-MO mdx 4cv mice. Scale bars = 50 μm. ( B , C , D ) Quantification of minimum muscle fiber diameter reveals a significant increase in myofiber size in THI-treated animals. Increased myofiber diameter was observed in both ( B ) injured and ( C ) uninjured quadriceps from THI-treated 11-MO mdx 4cv mice, whereas only ( D ) uninjured quadriceps in THI-treated 16-MO mdx 4cv mice showed increased myofiber size compared to vehicle controls. As indicated by the distributions, mean and median values of muscle fiber minimum diameters, there is an overall increase in muscle fiber size with THI treatment. Quantifications were undertaken in random fields in both injured and uninjured muscles in order to obtain an overall representation of fiber size increase for each muscle.* P <0.05, *** P <0.0005 by student’s t -test. Error bars represent SEM. DAPI, 4',6-diamidino-2-phenylindole; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Staining

    S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: S1P promotes functional improvement of mdx ( C57BL/10ScSn-Dmd mdx/J ) muscle. ( A ) Experimental schematic of longer-term, 14-day treatment of THI or PBS (vehicle) following CTX injury. THI was administered following the aforementioned dose and injection regimen. Following treatment, EDL muscles were harvested and specific isometric force was analyzed by in vitro myography from both injured and uninjured limbs. ( B ) Force frequency analysis reveals that EDL muscles isolated from injured limbs of THI-treated animals (n = 10) have significantly greater specific force compared to injured vehicle controls (n = 9). ( C ) Analysis of untreated and uninjured wt ( C57BL/10ScSn ) and mdx ( C57BL/10ScSn-Dmd mdx/J ) indicate specific force improved in injured but not uninjured THI-treated EDL muscles. ( D ) Incubation of uninjured and untreated mdx ( C57BL/10ScSn-Dmd mdx/J ) EDL muscles with a high concentration of S1P (10 μM) leads to a significant increase in maximal specific force. * P <0.05, ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; EDL, extensor digitorum longus; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Functional Assay, Injection, In Vitro, Isolation, Incubation, Concentration Assay

    Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: Direct administration of S1P promotes muscle regeneration following acute injury. ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of 3-MO mdx 4cv :Myf5 nlacZ/+ mice (n = 3, left TAs injected S1P, right TAs injected PBS) following CTX injury. ( B ) Top row: X-gal staining reveals an increased number of β-galactosidase+ nuclei at the sites of injury in S1P-treated TA muscles compared to vehicle controls. Bottom row: staining for eMyHC with DAB reveals a significant increase in the number of newly regenerated muscle fibers in S1P-treated TA muscles. Scale bars = 50 μm. ( C ) Left graph: quantification of β-galactosidase+ nuclei indicates the number of Myf5 + cells is significantly increased at the site of injury in S1P-treated compared to untreated muscles. Middle graph: a significant increase in β-galactosidase+ nuclei was also observed over the entire CSA of each S1P-treated TA muscle. Right graph: quantification of the number of eMyHC fibers within areas of regeneration was significantly greater with S1P treatment. * P <0.05 by student’s t -test. Error bars represent SEM. CSA, cross-sectional area; CTX, cardiotoxin; DAB, 3,3'-diaminobenzidine; eMyHC, embryonic myosin heavy chain; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Injection, Staining

    Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: Administration of S1P leads to increased levels of S1PR1 and P-rpS6 in vivo . ( A ) Experimental schematic of S1P and PBS (vehicle) injected daily for the first 72 hours into TAs of uninjured mdx 4cv mice (n = 4, 2.5-MO, left TAs injected S1P, right TAs injected PBS). ( B ) Western blot analysis of injected TAs (n = 3, 2.5-MO mdx 4cv ) indicates that administration of S1P significantly increases S1PR1 levels. ( C ) Western blot analysis of injected TAs (n = 4, 2.5-MO mdx 4cv ) for total, and P-Akt, P-mTOR and P-rpS6, reveals that total and P-rpS6 were significantly higher with S1P treatment. Increased levels of total and P-rpS6 suggest that S1P administration promotes protein synthesis in mdx muscles. * P <0.05 by student’s t -test. Error bars represent SEM. MO, month-old; P-Akt, phosphorylated Akt; P-mTOR, phosphorylated mammalian target of rapamycin; P-rpS6, phosphorylated ribosomal protein S6; rpS6, ribosomal protein S6; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; TA, tibialis anterior.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: In Vivo, Injection, Western Blot

    Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: Direct injection results in elevated S1P levels which correlate with the activation of receptor 1 in muscle fibers. ( A ) To quantify the elevation of S1P following direct administration, we injected a single dose (same dose as Figure ) of S1P in left TAs and vehicle in right TAs of uninjured mdx 4cv (n = 3, 11-MO) mice. TA muscles were harvested 15 minutes post injection for analysis by LC-MS/MS. Results indicate a significant elevation of S1P following direct injection. ( B ) To visualize the location of S1P following injection, biotinylated-S1P was injected in left TAs versus vehicle in right TAs of uninjured mdx 4cv mice (n = 2, 11-MO). Once more, TAs were harvested 15 minutes following injection. Staining with streptavidin conjugated to Alexa Fluor 594 reveals the presence of S1P-biotin around the perimeter of muscle fibers. ( C ) Staining of mdx 4cv TAs for S1PR1 and S1PR3 reveals S1PR1 is localized to the perimeter and perinuclear area (arrow) of muscle fibers (left photo). In contrast, staining for S1PR3 was mainly localized to the muscle vasculature (middle photo). Staining in parallel with an IgG isotype control for both antibodies shows the absence of non-specific staining (right graph). ( D ) Staining for S1PR1 in CTX-injured TAs (same tissue from Figure ) reveals S1PR1 is present at the perimeter and perinuclear area of regenerating eMyHC+ fibers. ( E ) Staining for phosphorylated S1PR1 in the same mdx 4cv TAs was more prominent in the perinuclear area of eMyHC+ fibers, indicating the presence of active S1PR1 signaling in regenerating fibers. Scale bars = 50 μm. ** P <0.005 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; eMyHC, embryonic myosin heavy chain; IgG, immunoglobulin G; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; S1PR3, S1P receptor 3; SEM, standard error of the mean; TA, tibialis anterior.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Injection, Activation Assay, Liquid Chromatography with Mass Spectroscopy, Staining, Liquid Chromatography, Mass Spectrometry

    IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: IP injection of THI reduces peripheral blood leukocytes and increases S1P levels in most tissues. ( A ) Leukocytes were analyzed from the peripheral blood of 1.5-MO mdx 4cv mice (n = 3) before and 12 hours following treatment with THI (2 × 250 μl 0.15 mg/ml IP injections, 6 hours apart). IP administration of THI significantly reduced circulating leukocytes to values below or near age-matched wt (n = 4). The average value of each population is listed in the table below the bar graph. Values between pre and post THI, and wt were also significant by ANOVA ( P <0.05) for all leukocytes except monocytes. ( B ) mdx 4cv mice (n = 6, 5-MO) were treated with THI or vehicle for 3 days (2 × 250 μl 0.15 mg/ml IP injections per day) following CTX injury to assess changes in S1P muscle content. Muscles and spleens were harvested on day 4 post injury for S1P analysis by LC-MS/MS. Results indicate S1P levels in spleen and injured quadriceps (quads) were significantly elevated with THI treatment. Interestingly, uninjured quadriceps did not show a significant increase of S1P, whereas uninjured TA muscles did. * P <0.05 by student’s t -test. Error bars represent SEM. CTX, cardiotoxin; IP, intraperitoneal; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MO, month-old; S1P, sphingosine-1-phoshate; SEM, standard error of the mean; TA, tibialis anterior; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole; wt, wild type.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Injection, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry

    Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Journal: Skeletal Muscle

    Article Title: Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice

    doi: 10.1186/2044-5040-3-20

    Figure Lengend Snippet: Longer-term treatment with THI elevated muscle force in uninjured mdx EDL muscles. ( A ) Experimental schematic outlining the treatment regimen. Beginning at 4 weeks of age, mdx 4cv mice (1-MO males) were treated for 4 weeks ad libitum with 50 mg/l THI (n = 4) or vehicle (n = 3) in drinking water. ( B ) Myography analysis of EDL muscles reveals a significant increase in maximal specific force with THI treatment. * P <0.05 by student’s t -test. Error bars represent SEM. ( C ) Summary of findings: S1P can act to not only promote myogenic cell activation and muscle repair, but also enhance muscle fiber size and force, possibly through S1PR1 mediated signaling. EDL, extensor digitorum longus; MO, month-old; S1P, sphingosine-1-phoshate; S1PR1, S1P receptor 1; SEM, standard error of the mean; THI, 2-acetyl-4(5)-tetrahydroxybutyl imidazole.

    Article Snippet: For injection of biotinylated-S1P, TAs from 11-MO mdx 4cv (n = 2) were injected intramuscularly with 20 μl 500 μM S1P-biotin or vehicle (Echelon Biosciences, Salt Lake City, UT, USA).

    Techniques: Activation Assay