s1pr1  (Abcam)

 
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    Name:
    Human EDG1 peptide
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    Catalog Number:
    AB39763
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    Structured Review

    Abcam s1pr1
    <t>S1PR1</t> expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

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    Images

    1) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    2) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    3) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    4) Product Images from "Conjugated bile acids promote cholangiocarcinoma cell invasive growth through activation of sphingosine 1-phosphate receptor 2"

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

    Journal: Hepatology (Baltimore, Md.)

    doi: 10.1002/hep.27085

    Differential expression of S1PRs in CCA cells and human CCA tissue. Total cell lysates of (A) human HuCCT1, CCLP1, and SG231 cells and (B) rat BDE1, BDEsp-TDE H10, and BDEsp-TDF E4 cells were prepared as previously described. 34 Protein levels of S1PR1, S1PR2, and S1PR3 were determined by western blotting analysis using specific Abs. β-actin was used as loading control. Representative images are shown. (C) Fluorescent IHC staining of S1PR2 in human CCA tissues. Human CCA tumor tissue and nontumor tissue from the same patient were processed for fluorescent IHC staining of S1PR2, as described in Materials and Methods. Representative images are shown. (a) Negative control (NC) without primary and second Ab. (b) NC without primary Ab. (c and d) Nontumor tissues stained with S1PR2. (e and f) Tumor tissues stained with S1PR2.
    Figure Legend Snippet: Differential expression of S1PRs in CCA cells and human CCA tissue. Total cell lysates of (A) human HuCCT1, CCLP1, and SG231 cells and (B) rat BDE1, BDEsp-TDE H10, and BDEsp-TDF E4 cells were prepared as previously described. 34 Protein levels of S1PR1, S1PR2, and S1PR3 were determined by western blotting analysis using specific Abs. β-actin was used as loading control. Representative images are shown. (C) Fluorescent IHC staining of S1PR2 in human CCA tissues. Human CCA tumor tissue and nontumor tissue from the same patient were processed for fluorescent IHC staining of S1PR2, as described in Materials and Methods. Representative images are shown. (a) Negative control (NC) without primary and second Ab. (b) NC without primary Ab. (c and d) Nontumor tissues stained with S1PR2. (e and f) Tumor tissues stained with S1PR2.

    Techniques Used: Expressing, Western Blot, Immunohistochemistry, Staining, Negative Control

    Differential expression of S1PRs in CCA cells. Total cellular RNA was isolated from (A) rat BDE1, (B) rat BDEsp, (C) rat BDEsp-TDE H10, (D) human HuCCT1, (E) CCLP1, and (F) SG231cells. mRNA levels of individual S1PRs were detected by real-time RT-PCR, as described in Materials and Methods, and normalized using β-actin or GAPDH as an internal control. Relative mRNA levels of S1PR2 and S1PR3 to S1PR1 (designated = 1) are shown. *** P
    Figure Legend Snippet: Differential expression of S1PRs in CCA cells. Total cellular RNA was isolated from (A) rat BDE1, (B) rat BDEsp, (C) rat BDEsp-TDE H10, (D) human HuCCT1, (E) CCLP1, and (F) SG231cells. mRNA levels of individual S1PRs were detected by real-time RT-PCR, as described in Materials and Methods, and normalized using β-actin or GAPDH as an internal control. Relative mRNA levels of S1PR2 and S1PR3 to S1PR1 (designated = 1) are shown. *** P

    Techniques Used: Expressing, Isolation, Quantitative RT-PCR

    5) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    6) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    7) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    8) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    9) Product Images from "Non-negligible factors in studying the ApoM-S1P axis using EA.hy926 cells"

    Article Title: Non-negligible factors in studying the ApoM-S1P axis using EA.hy926 cells

    Journal: Annals of Translational Medicine

    doi: 10.21037/atm.2020.03.74

    Immunofluorescence staining of S1PR1, S1PR2, S1PR3, S1PR4, and S1PR5 in EA.hy926 cells. S1PR1–5 antibodies were incubated at 4 °C overnight. FITC-conjugated secondary antibody was incubated at 37 °C for 1 h in the dark. Green indicates positive staining. S1PR1–3 expression was detected in EA.hy926 cells, whereas S1PR4 and S1PR5 were barely detected. S1PR1 was highly expressed and showed a dense patchy distribution, whereas S1PR2 and S1PR3 were expressed at low levels and showed a low-density punctate distribution. Magnification, ×200, scale bar =50 µm.
    Figure Legend Snippet: Immunofluorescence staining of S1PR1, S1PR2, S1PR3, S1PR4, and S1PR5 in EA.hy926 cells. S1PR1–5 antibodies were incubated at 4 °C overnight. FITC-conjugated secondary antibody was incubated at 37 °C for 1 h in the dark. Green indicates positive staining. S1PR1–3 expression was detected in EA.hy926 cells, whereas S1PR4 and S1PR5 were barely detected. S1PR1 was highly expressed and showed a dense patchy distribution, whereas S1PR2 and S1PR3 were expressed at low levels and showed a low-density punctate distribution. Magnification, ×200, scale bar =50 µm.

    Techniques Used: Immunofluorescence, Staining, Incubation, Expressing

    S1PRs mRNA and protein expression in EA.hy926 cells. The amplification curves of human S1PR1–5/GAPDH detected by duplex reverse transcription quantitative polymerase reaction. (A) The amplification curves of S1PR1–5 for 40 cycles (FAM channel, 465–510 nm); (B) the amplification curves of S1PR2 for 50 cycles (FAM channel, 465–510 nm); (C) the amplification curves of GAPDH for 40 cycles (CY5 channel, 618–660 nm); (D) the amplification curves of GAPDH for 50 cycles (CY5 channel, 618–660 nm); (E) the protein mass of S1PR1–5 in EA.hy926 cells. Three wells were loaded for each target protein, and the protein loading for each well was 60, 30, and 15 µg.
    Figure Legend Snippet: S1PRs mRNA and protein expression in EA.hy926 cells. The amplification curves of human S1PR1–5/GAPDH detected by duplex reverse transcription quantitative polymerase reaction. (A) The amplification curves of S1PR1–5 for 40 cycles (FAM channel, 465–510 nm); (B) the amplification curves of S1PR2 for 50 cycles (FAM channel, 465–510 nm); (C) the amplification curves of GAPDH for 40 cycles (CY5 channel, 618–660 nm); (D) the amplification curves of GAPDH for 50 cycles (CY5 channel, 618–660 nm); (E) the protein mass of S1PR1–5 in EA.hy926 cells. Three wells were loaded for each target protein, and the protein loading for each well was 60, 30, and 15 µg.

    Techniques Used: Expressing, Amplification

    The effects of ApoM overexpression on S1PR1–3 mRNA and protein levels in EA.hy926 cells. (A) S1PR1 mRNA levels in the OE group compared with the NC group. **, P
    Figure Legend Snippet: The effects of ApoM overexpression on S1PR1–3 mRNA and protein levels in EA.hy926 cells. (A) S1PR1 mRNA levels in the OE group compared with the NC group. **, P

    Techniques Used: Over Expression

    10) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    11) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    12) Product Images from "Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats"

    Article Title: Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats

    Journal: EXCLI Journal

    doi:

    Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P
    Figure Legend Snippet: Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P

    Techniques Used: Expressing, Injection, Western Blot

    13) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    14) Product Images from "Filamin A Expression Negatively Regulates Sphingosine-1-Phosphate-Induced NF-κB Activation in Melanoma Cells by Inhibition of Akt Signaling"

    Article Title: Filamin A Expression Negatively Regulates Sphingosine-1-Phosphate-Induced NF-κB Activation in Melanoma Cells by Inhibition of Akt Signaling

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00554-15

    S1P activates NF-κB via S1PR1 and S1PR2. (A and B) M2 and A7 melanoma cells were serum starved overnight. (A) RNA was isolated, and levels of S1PR1 to -3 mRNAs were quantified by QPCR and normalized to GAPDH levels. Data are expressed as relative
    Figure Legend Snippet: S1P activates NF-κB via S1PR1 and S1PR2. (A and B) M2 and A7 melanoma cells were serum starved overnight. (A) RNA was isolated, and levels of S1PR1 to -3 mRNAs were quantified by QPCR and normalized to GAPDH levels. Data are expressed as relative

    Techniques Used: Isolation, Real-time Polymerase Chain Reaction

    15) Product Images from "Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway"

    Article Title: Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway

    Journal: Nutrients

    doi: 10.3390/nu9030288

    Effect of treatment with protocatechuic acid (PCA) on SphK/S1P signaling pathway-related genes in mice with 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis. Representative sections of SphK1 immunohistochemistry in colonic tissues from C, C + PCA, TNBS, TNBS + PCA30 and TNBS + PCA60, and quantification of the positive cells. Image analysis was performed using the ImageJ software v3.91 with ten non-overlapping randomly chosen histological fields. Photographs are taken under magnification ×200 ( A ). Representative blots for SphK1 and S1PR1 protein, and results of densitometric quantification ( B ). mRNA expressions of SphK1, S1PR1, S1PL and SGPP2 genes ( C ), and level of S1P in colon homogenates analyzed by ELISA ( D ). Values are expressed as the means ± SEM of six mice per group. a p
    Figure Legend Snippet: Effect of treatment with protocatechuic acid (PCA) on SphK/S1P signaling pathway-related genes in mice with 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis. Representative sections of SphK1 immunohistochemistry in colonic tissues from C, C + PCA, TNBS, TNBS + PCA30 and TNBS + PCA60, and quantification of the positive cells. Image analysis was performed using the ImageJ software v3.91 with ten non-overlapping randomly chosen histological fields. Photographs are taken under magnification ×200 ( A ). Representative blots for SphK1 and S1PR1 protein, and results of densitometric quantification ( B ). mRNA expressions of SphK1, S1PR1, S1PL and SGPP2 genes ( C ), and level of S1P in colon homogenates analyzed by ELISA ( D ). Values are expressed as the means ± SEM of six mice per group. a p

    Techniques Used: Mouse Assay, Immunohistochemistry, Software, Enzyme-linked Immunosorbent Assay

    16) Product Images from "Targeting S1PR1/STAT3 loop abrogates desmoplasia and chemosensitizes pancreatic cancer to gemcitabine"

    Article Title: Targeting S1PR1/STAT3 loop abrogates desmoplasia and chemosensitizes pancreatic cancer to gemcitabine

    Journal: Theranostics

    doi: 10.7150/thno.25308

    FTY720 enhanced the effects of gemcitabine in pancreatic cancer cell lines. (A) MIA PaCa-2 cells were treated with indicating concentrations of FTY720and gemcitabine (for 24h) and cell survival was analyzed using MTT assay. (B) MIA PaCa-2 cells were treated with FTY720 in combination with gemcitabine and the loss in mitochondrial membrane potential was quantified using DiCO6(3) staining after 24h. (C) The clonogenic potential of the cells was quantified after 24 h pre-treatment of cells with the drug alone and the combinations and incubating for 24h. Cells were collected and plated again as 1000 cells per well and allowed them to grow in drug free environment. Representative images were shown in left panel. Then the colonies were fixed with 10% neutral buffered formalin and stained with crystal violet. The quantification was performed using TECAN spectrophotometer at 590nm after melting the stained colonies (right panel). (D) MIA PaCa-2 cells were treated with FTY720 and in combination with gemcitabine for 24h and the expression of S1PR1, p-STAT3, STAT3, and c-Myc; EMT markers, E-cadherin, N-cadherin and Vimentin and proliferative markers CyclinD1 and Cox-2 expression were analyzed using western blotting.β-Tubulin served as internal control
    Figure Legend Snippet: FTY720 enhanced the effects of gemcitabine in pancreatic cancer cell lines. (A) MIA PaCa-2 cells were treated with indicating concentrations of FTY720and gemcitabine (for 24h) and cell survival was analyzed using MTT assay. (B) MIA PaCa-2 cells were treated with FTY720 in combination with gemcitabine and the loss in mitochondrial membrane potential was quantified using DiCO6(3) staining after 24h. (C) The clonogenic potential of the cells was quantified after 24 h pre-treatment of cells with the drug alone and the combinations and incubating for 24h. Cells were collected and plated again as 1000 cells per well and allowed them to grow in drug free environment. Representative images were shown in left panel. Then the colonies were fixed with 10% neutral buffered formalin and stained with crystal violet. The quantification was performed using TECAN spectrophotometer at 590nm after melting the stained colonies (right panel). (D) MIA PaCa-2 cells were treated with FTY720 and in combination with gemcitabine for 24h and the expression of S1PR1, p-STAT3, STAT3, and c-Myc; EMT markers, E-cadherin, N-cadherin and Vimentin and proliferative markers CyclinD1 and Cox-2 expression were analyzed using western blotting.β-Tubulin served as internal control

    Techniques Used: MTT Assay, Staining, Spectrophotometry, Expressing, Western Blot

    FTY720 in co-operation with gemcitabine inhibited cell proliferation and downregulated the activation of NF-kB and S1PR1/STAT3 pathways and PP2A activation in pancreatic cancer. (A) Immunohistochemical analysis of proliferative markers cyclin D1, β-catenin and Ki-67 in tissue samples (upper panel). The quantification positive cells are shown lower panel. (B) FTY treatment inhibited the activation of NF-κB and NF-κB dependent gene expression. Immunohistochemical analysis of nuclear translocation of p65, representative image (left panel) and quantification of nuclear positive cells (right panel). Statistical analysis was performed using one way ANOVA followed by post hoc Tukey test; * p≤0.001 vs control. (C) The circulatory TNF in serum was measured using ELISA. Statistical significance was calculated using t -test, * p≤0.001 vs control. (D) PP2A enzyme activity was measured using a commercially available kit. Statistical significance was calculated using t -test, ** p≤0.05 vs control. (E) FTY720 inhibited the S1PR1/STAT3 loop and downstream signaling in pancreatic cancer. Immunoblot analysis showing the expression of indicated proteins in tissue lysates of pancreatic cancer samples.
    Figure Legend Snippet: FTY720 in co-operation with gemcitabine inhibited cell proliferation and downregulated the activation of NF-kB and S1PR1/STAT3 pathways and PP2A activation in pancreatic cancer. (A) Immunohistochemical analysis of proliferative markers cyclin D1, β-catenin and Ki-67 in tissue samples (upper panel). The quantification positive cells are shown lower panel. (B) FTY treatment inhibited the activation of NF-κB and NF-κB dependent gene expression. Immunohistochemical analysis of nuclear translocation of p65, representative image (left panel) and quantification of nuclear positive cells (right panel). Statistical analysis was performed using one way ANOVA followed by post hoc Tukey test; * p≤0.001 vs control. (C) The circulatory TNF in serum was measured using ELISA. Statistical significance was calculated using t -test, * p≤0.001 vs control. (D) PP2A enzyme activity was measured using a commercially available kit. Statistical significance was calculated using t -test, ** p≤0.05 vs control. (E) FTY720 inhibited the S1PR1/STAT3 loop and downstream signaling in pancreatic cancer. Immunoblot analysis showing the expression of indicated proteins in tissue lysates of pancreatic cancer samples.

    Techniques Used: Activation Assay, Immunohistochemistry, Expressing, Translocation Assay, Enzyme-linked Immunosorbent Assay, Activity Assay

    17) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    18) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    19) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    20) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    21) Product Images from "Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats"

    Article Title: Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats

    Journal: EXCLI Journal

    doi:

    Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P
    Figure Legend Snippet: Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P

    Techniques Used: Expressing, Injection, Western Blot

    22) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    23) Product Images from "Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats"

    Article Title: Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats

    Journal: EXCLI Journal

    doi:

    Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P
    Figure Legend Snippet: Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P

    Techniques Used: Expressing, Injection, Western Blot

    24) Product Images from "MicroRNA-708-5p acts as a therapeutic agent against metastatic lung cancer"

    Article Title: MicroRNA-708-5p acts as a therapeutic agent against metastatic lung cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.6594

    p21 is a direct target of miR-708-5p ( A ) Immunoblotting for endogenous p21 and S1PR1 proteins in the A549, H1299 and PG cells transfected with the miR-708-5p or control mimics. β-actin was used as a loading control. ( B ) Luciferase activity of the wild-type or mutant p21 3′UTR reporter genes in the 293T cells transfected with the miR-708-5p or control mimics. ( C ) Immunoblotting of p21 in the 95C and QG56 cells transfected with antago-708-5p or control. β-actin was used as a loading control. ( D – F ) Matrigel invasion and apoptosis assay in A549 cells co-transfected with p21 expressing vector or control vector and miR-708-5p or control mimics. ( G – I ) Matrigel invasion and apoptosis in the 95C cells co-transfected with antago-708-5p or antago control and p21 siRNA (si-p21) or control siRNA (si-control). (D, G) Upper: inversion imagines of the cells penetrating through the matrigel after stained with 0.25% Trypan Blue. Scale bars, 250 μm. Lower: means and s.d. of the number of the cells penetrating through the transwell membrane. (E, H) Immunoblotting of active caspase-3 and cleaved PARP in A549 (E) and 95C (H). (F, I) means and s.d. of fractions of apoptotic cells (early and late apoptosis) detected using Annexin V/PI kit in A549 (F) and 95C (I) cells.
    Figure Legend Snippet: p21 is a direct target of miR-708-5p ( A ) Immunoblotting for endogenous p21 and S1PR1 proteins in the A549, H1299 and PG cells transfected with the miR-708-5p or control mimics. β-actin was used as a loading control. ( B ) Luciferase activity of the wild-type or mutant p21 3′UTR reporter genes in the 293T cells transfected with the miR-708-5p or control mimics. ( C ) Immunoblotting of p21 in the 95C and QG56 cells transfected with antago-708-5p or control. β-actin was used as a loading control. ( D – F ) Matrigel invasion and apoptosis assay in A549 cells co-transfected with p21 expressing vector or control vector and miR-708-5p or control mimics. ( G – I ) Matrigel invasion and apoptosis in the 95C cells co-transfected with antago-708-5p or antago control and p21 siRNA (si-p21) or control siRNA (si-control). (D, G) Upper: inversion imagines of the cells penetrating through the matrigel after stained with 0.25% Trypan Blue. Scale bars, 250 μm. Lower: means and s.d. of the number of the cells penetrating through the transwell membrane. (E, H) Immunoblotting of active caspase-3 and cleaved PARP in A549 (E) and 95C (H). (F, I) means and s.d. of fractions of apoptotic cells (early and late apoptosis) detected using Annexin V/PI kit in A549 (F) and 95C (I) cells.

    Techniques Used: Transfection, Luciferase, Activity Assay, Mutagenesis, Apoptosis Assay, Expressing, Plasmid Preparation, Staining

    25) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    26) Product Images from "The sphingosine-1-phosphate/RhoA/Rho associated kinases/myosin light chain pathway in detrusor of female rats is down-regulated in response to ovariectomy"

    Article Title: The sphingosine-1-phosphate/RhoA/Rho associated kinases/myosin light chain pathway in detrusor of female rats is down-regulated in response to ovariectomy

    Journal: Chinese Medical Journal

    doi: 10.1097/CM9.0000000000000767

    Relative expression of protein in rats detrusor by Western blotting. (A) Relative expression of SphK1/2 protein. (B) Relative expression of S1PR1/2/3 protein. (C) Relative expression of RhoA, ROCK1 and ROCK2 protein. (D) Relative expression of p-MYPT1, MYPT1, p-MLC20, and MLC20 protein. ∗ P
    Figure Legend Snippet: Relative expression of protein in rats detrusor by Western blotting. (A) Relative expression of SphK1/2 protein. (B) Relative expression of S1PR1/2/3 protein. (C) Relative expression of RhoA, ROCK1 and ROCK2 protein. (D) Relative expression of p-MYPT1, MYPT1, p-MLC20, and MLC20 protein. ∗ P

    Techniques Used: Expressing, Western Blot

    Relative expression of mRNA in rats detrusor by quantitative real-time polymerase chain reaction. (A) Relative expression of SphK1/2 mRNA. (B) Relative expression of S1PR1/2/3 mRNA. (C) Relative expression of RhoA and ROCK1/2 mRNA. (D) Relative expression of MYPT1 and MLC20 mRNA. ∗ P
    Figure Legend Snippet: Relative expression of mRNA in rats detrusor by quantitative real-time polymerase chain reaction. (A) Relative expression of SphK1/2 mRNA. (B) Relative expression of S1PR1/2/3 mRNA. (C) Relative expression of RhoA and ROCK1/2 mRNA. (D) Relative expression of MYPT1 and MLC20 mRNA. ∗ P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    27) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    28) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    29) Product Images from "Targeting S1PR1/STAT3 loop abrogates desmoplasia and chemosensitizes pancreatic cancer to gemcitabine"

    Article Title: Targeting S1PR1/STAT3 loop abrogates desmoplasia and chemosensitizes pancreatic cancer to gemcitabine

    Journal: Theranostics

    doi: 10.7150/thno.25308

    FTY720 enhanced the effects of gemcitabine in pancreatic cancer cell lines. (A) MIA PaCa-2 cells were treated with indicating concentrations of FTY720and gemcitabine (for 24h) and cell survival was analyzed using MTT assay. (B) MIA PaCa-2 cells were treated with FTY720 in combination with gemcitabine and the loss in mitochondrial membrane potential was quantified using DiCO6(3) staining after 24h. (C) The clonogenic potential of the cells was quantified after 24 h pre-treatment of cells with the drug alone and the combinations and incubating for 24h. Cells were collected and plated again as 1000 cells per well and allowed them to grow in drug free environment. Representative images were shown in left panel. Then the colonies were fixed with 10% neutral buffered formalin and stained with crystal violet. The quantification was performed using TECAN spectrophotometer at 590nm after melting the stained colonies (right panel). (D) MIA PaCa-2 cells were treated with FTY720 and in combination with gemcitabine for 24h and the expression of S1PR1, p-STAT3, STAT3, and c-Myc; EMT markers, E-cadherin, N-cadherin and Vimentin and proliferative markers CyclinD1 and Cox-2 expression were analyzed using western blotting.β-Tubulin served as internal control
    Figure Legend Snippet: FTY720 enhanced the effects of gemcitabine in pancreatic cancer cell lines. (A) MIA PaCa-2 cells were treated with indicating concentrations of FTY720and gemcitabine (for 24h) and cell survival was analyzed using MTT assay. (B) MIA PaCa-2 cells were treated with FTY720 in combination with gemcitabine and the loss in mitochondrial membrane potential was quantified using DiCO6(3) staining after 24h. (C) The clonogenic potential of the cells was quantified after 24 h pre-treatment of cells with the drug alone and the combinations and incubating for 24h. Cells were collected and plated again as 1000 cells per well and allowed them to grow in drug free environment. Representative images were shown in left panel. Then the colonies were fixed with 10% neutral buffered formalin and stained with crystal violet. The quantification was performed using TECAN spectrophotometer at 590nm after melting the stained colonies (right panel). (D) MIA PaCa-2 cells were treated with FTY720 and in combination with gemcitabine for 24h and the expression of S1PR1, p-STAT3, STAT3, and c-Myc; EMT markers, E-cadherin, N-cadherin and Vimentin and proliferative markers CyclinD1 and Cox-2 expression were analyzed using western blotting.β-Tubulin served as internal control

    Techniques Used: MTT Assay, Staining, Spectrophotometry, Expressing, Western Blot

    FTY720 in co-operation with gemcitabine inhibited cell proliferation and downregulated the activation of NF-kB and S1PR1/STAT3 pathways and PP2A activation in pancreatic cancer. (A) Immunohistochemical analysis of proliferative markers cyclin D1, β-catenin and Ki-67 in tissue samples (upper panel). The quantification positive cells are shown lower panel. (B) FTY treatment inhibited the activation of NF-κB and NF-κB dependent gene expression. Immunohistochemical analysis of nuclear translocation of p65, representative image (left panel) and quantification of nuclear positive cells (right panel). Statistical analysis was performed using one way ANOVA followed by post hoc Tukey test; * p≤0.001 vs control. (C) The circulatory TNF in serum was measured using ELISA. Statistical significance was calculated using t -test, * p≤0.001 vs control. (D) PP2A enzyme activity was measured using a commercially available kit. Statistical significance was calculated using t -test, ** p≤0.05 vs control. (E) FTY720 inhibited the S1PR1/STAT3 loop and downstream signaling in pancreatic cancer. Immunoblot analysis showing the expression of indicated proteins in tissue lysates of pancreatic cancer samples.
    Figure Legend Snippet: FTY720 in co-operation with gemcitabine inhibited cell proliferation and downregulated the activation of NF-kB and S1PR1/STAT3 pathways and PP2A activation in pancreatic cancer. (A) Immunohistochemical analysis of proliferative markers cyclin D1, β-catenin and Ki-67 in tissue samples (upper panel). The quantification positive cells are shown lower panel. (B) FTY treatment inhibited the activation of NF-κB and NF-κB dependent gene expression. Immunohistochemical analysis of nuclear translocation of p65, representative image (left panel) and quantification of nuclear positive cells (right panel). Statistical analysis was performed using one way ANOVA followed by post hoc Tukey test; * p≤0.001 vs control. (C) The circulatory TNF in serum was measured using ELISA. Statistical significance was calculated using t -test, * p≤0.001 vs control. (D) PP2A enzyme activity was measured using a commercially available kit. Statistical significance was calculated using t -test, ** p≤0.05 vs control. (E) FTY720 inhibited the S1PR1/STAT3 loop and downstream signaling in pancreatic cancer. Immunoblot analysis showing the expression of indicated proteins in tissue lysates of pancreatic cancer samples.

    Techniques Used: Activation Assay, Immunohistochemistry, Expressing, Translocation Assay, Enzyme-linked Immunosorbent Assay, Activity Assay

    30) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    31) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    32) Product Images from "Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway"

    Article Title: Protective Effect of Protocatechuic Acid on TNBS-Induced Colitis in Mice Is Associated with Modulation of the SphK/S1P Signaling Pathway

    Journal: Nutrients

    doi: 10.3390/nu9030288

    Effect of treatment with protocatechuic acid (PCA) on SphK/S1P signaling pathway-related genes in mice with 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis. Representative sections of SphK1 immunohistochemistry in colonic tissues from C, C + PCA, TNBS, TNBS + PCA30 and TNBS + PCA60, and quantification of the positive cells. Image analysis was performed using the ImageJ software v3.91 with ten non-overlapping randomly chosen histological fields. Photographs are taken under magnification ×200 ( A ). Representative blots for SphK1 and S1PR1 protein, and results of densitometric quantification ( B ). mRNA expressions of SphK1, S1PR1, S1PL and SGPP2 genes ( C ), and level of S1P in colon homogenates analyzed by ELISA ( D ). Values are expressed as the means ± SEM of six mice per group. a p
    Figure Legend Snippet: Effect of treatment with protocatechuic acid (PCA) on SphK/S1P signaling pathway-related genes in mice with 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis. Representative sections of SphK1 immunohistochemistry in colonic tissues from C, C + PCA, TNBS, TNBS + PCA30 and TNBS + PCA60, and quantification of the positive cells. Image analysis was performed using the ImageJ software v3.91 with ten non-overlapping randomly chosen histological fields. Photographs are taken under magnification ×200 ( A ). Representative blots for SphK1 and S1PR1 protein, and results of densitometric quantification ( B ). mRNA expressions of SphK1, S1PR1, S1PL and SGPP2 genes ( C ), and level of S1P in colon homogenates analyzed by ELISA ( D ). Values are expressed as the means ± SEM of six mice per group. a p

    Techniques Used: Mouse Assay, Immunohistochemistry, Software, Enzyme-linked Immunosorbent Assay

    33) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    34) Product Images from "Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway"

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    Journal: United European Gastroenterology Journal

    doi: 10.1177/2050640616637968

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage
    Figure Legend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Techniques Used: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased
    Figure Legend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Techniques Used: Western Blot, Expressing

    35) Product Images from "Sphingosine-1-Phosphate Enhances Satellite Cell Activation in Dystrophic Muscles through a S1PR2/STAT3 Signaling Pathway"

    Article Title: Sphingosine-1-Phosphate Enhances Satellite Cell Activation in Dystrophic Muscles through a S1PR2/STAT3 Signaling Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0037218

    Dynamic changes in S1P signaling after muscle injury. A) Heat map of S1P receptors, S1P metabolic genes and other markers in WT C57BL/6 mouse skeletal muscle from day 0–40 after NTX injection. Red is induced, green is repressed relative to the mean. Note: two probes were available for some genes. Lox = lysyl oxidase; S1pr1–5 = S1PRs; Sgpl1 = SPL. Sgpp = S1P phosphatases; Sphk = sphingosine kinases. B) SPL protein (Sgpl1p) expression is induced after muscle injury, as determined by immunoblotting (above) and imageJ quantification relative to day 0 (uninjured muscle) and normalized to actin (below). C) Relative gene expression of Sphk1, Sphk2, Sgpl1 and Sgpp1 (normalized to Gapdh ) were evaluated in whole muscle over time after injury by qRT-PCR. Gene levels are depicted relative to their own baseline levels (day 0 set as 1 for all genes). D) Corresponding plasma S1P levels were measured by LC-MS, E) Relative gene expression of S1PRs was determined as in “C” and is depicted relative to S1pr1 (day 0 set as 1 for S1pr1 only). F) S1PR induction is shown relative to baseline levels for each receptor (day 0 set as 1 for all genes). * indicates significant difference compared to day 0, p≤0.05. Data are means ± SD; n = 3–5/group.
    Figure Legend Snippet: Dynamic changes in S1P signaling after muscle injury. A) Heat map of S1P receptors, S1P metabolic genes and other markers in WT C57BL/6 mouse skeletal muscle from day 0–40 after NTX injection. Red is induced, green is repressed relative to the mean. Note: two probes were available for some genes. Lox = lysyl oxidase; S1pr1–5 = S1PRs; Sgpl1 = SPL. Sgpp = S1P phosphatases; Sphk = sphingosine kinases. B) SPL protein (Sgpl1p) expression is induced after muscle injury, as determined by immunoblotting (above) and imageJ quantification relative to day 0 (uninjured muscle) and normalized to actin (below). C) Relative gene expression of Sphk1, Sphk2, Sgpl1 and Sgpp1 (normalized to Gapdh ) were evaluated in whole muscle over time after injury by qRT-PCR. Gene levels are depicted relative to their own baseline levels (day 0 set as 1 for all genes). D) Corresponding plasma S1P levels were measured by LC-MS, E) Relative gene expression of S1PRs was determined as in “C” and is depicted relative to S1pr1 (day 0 set as 1 for S1pr1 only). F) S1PR induction is shown relative to baseline levels for each receptor (day 0 set as 1 for all genes). * indicates significant difference compared to day 0, p≤0.05. Data are means ± SD; n = 3–5/group.

    Techniques Used: Injection, Expressing, Quantitative RT-PCR, Liquid Chromatography with Mass Spectroscopy

    STAT3 and S1PR2 signaling interactions in muscle regeneration of WT and mdx mice. A) Mdx mice received NTX with or without THI treatment. Muscles were harvested on day 5 post-injury, and whole muscle extracts were examined for total STAT3 (STAT3-T) and phosphorylated STAT3 (STAT3-P) protein levels. Actin is used as a loading control. B) STAT3 activation in SC-derived myoblasts treated in vitro with 10 µM S1PR1 (R1i), S1PR2 (R2i), and S1PR3 (R3i) antagonists. S1PR1 is used as a loading control, as its expression is invariant under these conditions. C) WT SC-derived primary myoblasts were treated with S1PR2 antagonist (JTE-013) or vehicle, and whole cell extracts were examined for STAT3 phosphorylation, total STAT3 and p21 and p27 protein levels. Actin is used as a loading control. D) SC-derived primary myoblasts were treated with either 50 nM siRNA against S1PR2 or mock transfection. S1PR2 knockdown was confirmed by qRT-PCR. E) Whole cell extracts of control and S1PR2 knockdown cells were examined for phosphorylated STAT3 and actin protein levels by immunoblotting. p = 0.03 (siRNA vs. mock) for relative densitometric levels of STAT3-P/actin from two experiments performed in duplicates. F) WT mice were treated with NTX plus antagonist of S1PR2 (JTE-013) or vehicle by subcutaneous injection. Muscles were harvested 5 days post injury; regenerating fibers were imaged, and G) regenerating fibers were quantified. H) Flow cytometry analysis of SCs from WT mice treated with JTE-013 or vehicle at 5 days post injury; * indicates p≤0.05, data are means ± SD, n = 5/group.
    Figure Legend Snippet: STAT3 and S1PR2 signaling interactions in muscle regeneration of WT and mdx mice. A) Mdx mice received NTX with or without THI treatment. Muscles were harvested on day 5 post-injury, and whole muscle extracts were examined for total STAT3 (STAT3-T) and phosphorylated STAT3 (STAT3-P) protein levels. Actin is used as a loading control. B) STAT3 activation in SC-derived myoblasts treated in vitro with 10 µM S1PR1 (R1i), S1PR2 (R2i), and S1PR3 (R3i) antagonists. S1PR1 is used as a loading control, as its expression is invariant under these conditions. C) WT SC-derived primary myoblasts were treated with S1PR2 antagonist (JTE-013) or vehicle, and whole cell extracts were examined for STAT3 phosphorylation, total STAT3 and p21 and p27 protein levels. Actin is used as a loading control. D) SC-derived primary myoblasts were treated with either 50 nM siRNA against S1PR2 or mock transfection. S1PR2 knockdown was confirmed by qRT-PCR. E) Whole cell extracts of control and S1PR2 knockdown cells were examined for phosphorylated STAT3 and actin protein levels by immunoblotting. p = 0.03 (siRNA vs. mock) for relative densitometric levels of STAT3-P/actin from two experiments performed in duplicates. F) WT mice were treated with NTX plus antagonist of S1PR2 (JTE-013) or vehicle by subcutaneous injection. Muscles were harvested 5 days post injury; regenerating fibers were imaged, and G) regenerating fibers were quantified. H) Flow cytometry analysis of SCs from WT mice treated with JTE-013 or vehicle at 5 days post injury; * indicates p≤0.05, data are means ± SD, n = 5/group.

    Techniques Used: Mouse Assay, Activation Assay, Derivative Assay, In Vitro, Expressing, Transfection, Quantitative RT-PCR, Injection, Flow Cytometry, Cytometry

    36) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    37) Product Images from "Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats"

    Article Title: Activation of sphingosine 1-phosphate receptor-1 by SEW2871 improves cognitive function in Alzheimer's disease model rats

    Journal: EXCLI Journal

    doi:

    Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P
    Figure Legend Snippet: Effect of SEW2871 on S1PR1-3 expression in Aβ1-42 injected rats evaluated by Western blotting method. 15 days after the stereotaxic procedure, the expression of S1PR1-3 in hippocampus was detected. Results showed decreasing of S1PR1 (A) and increasing of S1PR2 (B) in AD model rats. Treatment of AD model rats with SEW2871 resulted in elevated levels of S1PR1 (A) in hippocampus and had no effect on S1P2 (B). The expression of S1P3 did not show any change between groups (C). β-actin protein was used here as an internal control. One representative Western blot is shown; n = 6. The band density values were calculated and the values from the control were used as 1. Values are the mean ± S.E.M., **P

    Techniques Used: Expressing, Injection, Western Blot

    38) Product Images from "Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells"

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1402304

    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    Figure Legend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.
    Figure Legend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Techniques Used: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.
    Figure Legend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Techniques Used: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p
    Figure Legend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    Related Articles

    other:

    Article Title: S1PR1 promotes proliferation and inhibits apoptosis of esophageal squamous cell carcinoma through activating STAT3 pathway
    Article Snippet: Previous studies have demonstrated that upregulation of S1PR1 was found in some solid human cancers, including breast cancer, gastric cancer and hepatocellular carcinoma (HCC) [ , – ].

    SDS Page:

    Article Title: A proof of concept phase I/II pilot trial of LSD1 inhibition by tranylcypromine combined with ATRA in refractory/relapsed AML patients not eligible for intensive therapy
    Article Snippet: Western blotCells were lysed in SDS lysis buffer (0,1% SDS, 50 mM Tris pH 8.0, 10U/ml Benzonase and Protease Inhibitor Cocktail (Sigma-Aldrich, Darmstadt Germany)) for 30 min at 4 °C. .. For target detection 0.5 µg total protein (for H3K4me2 and total H3 detection) and 5 µg total protein (for actin and LSD1 detection) were resolved on 12% Bis-Tris gels (Invitrogen) by SDS-PAGE, blotted onto nitrocellulose membranes, and probed by standard methods using the following antibodies: anti-β-actin (Sigma-Aldrich, A5441, 1:5000), anti-total H3 (Abcam, ab39763, 1:5000), anti-H3K4me2 (Abcam, ab32356, 1:5000), anti-LSD1 (Cell Signaling, # 2139S, 1:1000). ..

    Article Title: Loss of the Histone Methyltransferase EZH2 induces Resistance to Multiple Drugs in Acute Myeloid Leukemia
    Article Snippet: Cells were lysed in RIPA lysis buffer (50 mM Tris-HCL [pH 8], 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, protease inhibitors) for 30 min at 4°C. .. Protein extracts were resolved by SDS-PAGE and blotted to Nitrocellulose membranes and probed with the following antibodies: anti-beta Actin (Sigma-Aldrich, A5441), anti–EZH2 (AC22, Cell Signaling Technology, 3147S), anti-EZH2 (D2C9, Cell Signaling, 5246), anti-pT487 EZH2 (Abcam, ab109398), anti–SUZ12 (Cell Signaling Technology, 3737S), anti–EED (Millipore, 09-774), anti-EZH1 (Abcam, ab13665), anti-total H3 (Abcam, ab39763), anti-H3K27me3 (Cell Signaling Technologies, 9733), anti-V5 (Abcam, ab9116), anti-total CDK1 (Cell Signaling Technologies, 9112S), anti-pCDK1 (Cell Signaling Technologies, 9111S), anti-MRP1/ABCC1 (Santa Cruz, sc-18835), anti-Ubiquitin (Abcam, ab7780), anti-HOXB7 (Abcam, ab51237), anti-HOXA9 (Abcam, ab140631), anti-TRIM21 antibody (Abcam, ab91432), anti-HSP90 antibody (Abcam, ab13495) and anti-STIP1 antibody (Abcam, ab126724). .. Densitometric analysis was performed using ImageJ software (NIH, Bethesda, MD).

    Expressing:

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells
    Article Snippet: We found that S1PR1 expression spontaneously increased on normal B cells cultured in the absence of S1P ( , ), an effect that is inhibited when S1P is present ( ). .. Consistent with the results of others studying the effects of Ag receptor engagement on S1PR1 expression in mouse T cells , we found that spontaneous upregulation of S1PR1 on human B cells cultured in the absence of S1P is similarly repressed by BCR cross-linking ( ). .. Importantly, repression of S1PR1 expression by BCR cross-linking is reversed by pretreatment of normal B cells with idelalisib , suggesting a key role of PI3Kδ within the mechanism of this repression.

    Cell Culture:

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells
    Article Snippet: We found that S1PR1 expression spontaneously increased on normal B cells cultured in the absence of S1P ( , ), an effect that is inhibited when S1P is present ( ). .. Consistent with the results of others studying the effects of Ag receptor engagement on S1PR1 expression in mouse T cells , we found that spontaneous upregulation of S1PR1 on human B cells cultured in the absence of S1P is similarly repressed by BCR cross-linking ( ). .. Importantly, repression of S1PR1 expression by BCR cross-linking is reversed by pretreatment of normal B cells with idelalisib , suggesting a key role of PI3Kδ within the mechanism of this repression.

    Staining:

    Article Title: Moesin Controls Clathrin-Mediated S1PR1 Internalization in T Cells
    Article Snippet: Mouse CD4+ T cells were fixed, permeabilized, and stained with the anti-S1PR1 (5 µg/ml; ab11424) or control rabbit IgG followed by Alexa Fluor 488-conjugated anti-rabbit IgG. .. The cells were also stained with the anti-S1PR1 pre-absorbed with the antigen peptide (ab39763; Abcam) at a molar ratio of 1∶20 overnight. ..

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    <t>S1PR1</t> expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p
    S1pr1, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam antibody against s1pr1
    miR-363 inhibits the proliferation, migration and invasion of ccRCC cells by downregulating <t>S1PR1.</t> a Folds change of miR-363 and S1PR1 mRNA levels in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). b Folds change of miR-363 and S1PR1 mRNA levels in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). c MTS assay suggested that overexpression of S1PR1 reversed the negative proliferative effects of miR-363 mimics in 786O cells. d Knockdown of S1PR1 counteracted the positive proliferative effects of miR-363 inhibitor in 769P cells. e Transwell assays suggested that overexpression of S1PR1 reversed the negative migration and invasion effects of miR-363 mimics in 786O cells. f Knockdown of S1PR1 counteracted the positive migration and invasion effects of miR-363 inhibitor in 769P cells. g Alterations of S1PR1, ERK and downstream genes of ERK protein level in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). h Alterations of S1PR1, ERK and downstream genes of ERK protein level in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). Data are presented as the mean ± SD. (*P
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    S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Journal: The Journal of Immunology Author Choice

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    doi: 10.4049/jimmunol.1402304

    Figure Lengend Snippet: S1PR1 expression on normal and CLL B cells cultured in the absence of S1P. ( A ) Normal B cells from six healthy individuals were cultured for 16 h in medium lacking S1P and examined for S1PR1 expression by flow cytometry. An increase in S1PR1 was observed between 2 and 4 h and reached a peak at 8–16 h. ( B ) Normal B cells from six donors were cultured in S1P-free medium for 16 h in the presence or absence of anti-IgM and/or idelalisib (1 μM). S1PR1 expression was measured by flow cytometry. IgM cross-linking prevented the spontaneous increase in S1PR1 expression ( p = 0.0039). Idelalisib had little effect on S1PR1 expression on normal B cells in the absence of IgM ( p = 0.177); however, treatment reversed the anti-IgM–mediated suppression of S1PR1 expression ( p = 0.014) with levels returning to those of untreated cells at 16 h ( p = 0.177). ( C ) CLL cells from 20 patients were cultured for 16 h in S1P-free medium and examined for S1PR1 levels by flow cytometry. There was an overall increase in S1PR1 expression ( p

    Article Snippet: S1PR1 is underexpressed in normal germinal centers and CLL lymph nodes Expression of S1PR1 within human lymphoid tissues has hitherto not been described.

    Techniques: Expressing, Cell Culture, Flow Cytometry, Cytometry

    Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Journal: The Journal of Immunology Author Choice

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    doi: 10.4049/jimmunol.1402304

    Figure Lengend Snippet: Expression of S1PR1 in normal and CLL node. ( A and B ) Normal lymph node. The cells of the germinal center (GC) are identified by staining for CD23, which is mainly expressed on activated B cells. Staining of the adjacent section for S1PR1 shows that CD23 + cells do not express S1PR1, whereas the cells within the outer follicle (OF) stain positively. ( C – F ) Sequential section of CLL lymph nodes stained for S1PR1 and CD20. In (C) and (D), it can be clearly seen that the endothelial cells lining the sinus (S) and other endothelial cells express S1PR1, whereas CD20 + CLL cells lack the receptor. (E) and (F) show a representative proliferation center in a CLL lymph node. CD20 + CLL cells do not express S1PR1. Parallel staining with the relevant isotypic control Ab is shown in Supplemental Fig. 2A . Scale bar, 50 μM; original magnification ×20.

    Article Snippet: S1PR1 is underexpressed in normal germinal centers and CLL lymph nodes Expression of S1PR1 within human lymphoid tissues has hitherto not been described.

    Techniques: Expressing, Staining

    Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Journal: The Journal of Immunology Author Choice

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    doi: 10.4049/jimmunol.1402304

    Figure Lengend Snippet: Effect of BCR inhibitors on TEM toward S1P and CCL21. ( A ) CLL cells from five patients were incubated for 16 h in the presence or absence of idelalisib (1 μM) and then examined for migration toward S1P using HUVEC-coated transwells. The numbers above bars indicate mean fluorescence intensity values for S1PR1 staining at the end of the incubation period. Idelalisib increased TEM toward S1P in all cases, the amount of migration correlating with levels of S1PR1. Untreated CLL cells underwent little or no migration. ( B ) CLL cells from three patients were cultured in the absence or presence of idelalisib, fostamatinib, or ibrutinib (all at 1 μM) and examined for migration toward CCL21 using HUVEC-coated transwells. Untreated CLL cells underwent TEM toward CCL21. Migration was reduced by fostamatinib and ibrutinib, but not idelalisib.

    Article Snippet: S1PR1 is underexpressed in normal germinal centers and CLL lymph nodes Expression of S1PR1 within human lymphoid tissues has hitherto not been described.

    Techniques: Transmission Electron Microscopy, Incubation, Migration, Fluorescence, Staining, Cell Culture

    Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Journal: The Journal of Immunology Author Choice

    Article Title: Expression of Functional Sphingosine-1 Phosphate Receptor-1 Is Reduced by B Cell Receptor Signaling and Increased by Inhibition of PI3 Kinase δ but Not SYK or BTK in Chronic Lymphocytic Leukemia Cells

    doi: 10.4049/jimmunol.1402304

    Figure Lengend Snippet: Effect of idelalisib on S1PR1 expression in CLL cells. ( A ) CLL cells from 20 cases were cultured in the presence or absence of idelalisib (1 μM) and examined by flow cytometry for S1PR1 levels. The chart shows the fold increase in mean fluorescence intensity compared with cells cultured for the same amount of time in the absence of idelalisib. The increase in S1PR1 expression was statistically significant at both time points ( p

    Article Snippet: S1PR1 is underexpressed in normal germinal centers and CLL lymph nodes Expression of S1PR1 within human lymphoid tissues has hitherto not been described.

    Techniques: Expressing, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Journal: United European Gastroenterology Journal

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    doi: 10.1177/2050640616637968

    Figure Lengend Snippet: SphK1 inhibition improves hepatic hemorrhage in histopathology. Changes in hepatic histopathology were evaluated six hours after LPS/GalN administration with or without specific inhibitor for SphK1, S1PR1 or S1PR3. Quantification of hepatic hemorrhage

    Article Snippet: The phosphorylation of ERK was also reduced by S1PR1 or S1PR3 inhibition, but this was not significant ( ).

    Techniques: Inhibition, Histopathology

    Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Journal: United European Gastroenterology Journal

    Article Title: Sphingosine kinase 1 inhibition improves lipopolysaccharide/D-galactosamine-induced acute liver failure by inhibiting mitogen-activated protein kinases pathway

    doi: 10.1177/2050640616637968

    Figure Lengend Snippet: Protein levels of SphK1, S1PR1 and S1PR3 are upregulated by LPS/GalN. Western blotting was used to determine the expression of hepatic SphK1 (a), S1PR1 (b) and S1PR3 (c) after LPS/GalN administration. The expression of each protein was indeed decreased

    Article Snippet: The phosphorylation of ERK was also reduced by S1PR1 or S1PR3 inhibition, but this was not significant ( ).

    Techniques: Western Blot, Expressing

    miR-363 inhibits the proliferation, migration and invasion of ccRCC cells by downregulating S1PR1. a Folds change of miR-363 and S1PR1 mRNA levels in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). b Folds change of miR-363 and S1PR1 mRNA levels in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). c MTS assay suggested that overexpression of S1PR1 reversed the negative proliferative effects of miR-363 mimics in 786O cells. d Knockdown of S1PR1 counteracted the positive proliferative effects of miR-363 inhibitor in 769P cells. e Transwell assays suggested that overexpression of S1PR1 reversed the negative migration and invasion effects of miR-363 mimics in 786O cells. f Knockdown of S1PR1 counteracted the positive migration and invasion effects of miR-363 inhibitor in 769P cells. g Alterations of S1PR1, ERK and downstream genes of ERK protein level in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). h Alterations of S1PR1, ERK and downstream genes of ERK protein level in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). Data are presented as the mean ± SD. (*P

    Journal: Cancer Cell International

    Article Title: miR-363 suppresses the proliferation, migration and invasion of clear cell renal cell carcinoma by downregulating S1PR1

    doi: 10.1186/s12935-020-01313-9

    Figure Lengend Snippet: miR-363 inhibits the proliferation, migration and invasion of ccRCC cells by downregulating S1PR1. a Folds change of miR-363 and S1PR1 mRNA levels in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). b Folds change of miR-363 and S1PR1 mRNA levels in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). c MTS assay suggested that overexpression of S1PR1 reversed the negative proliferative effects of miR-363 mimics in 786O cells. d Knockdown of S1PR1 counteracted the positive proliferative effects of miR-363 inhibitor in 769P cells. e Transwell assays suggested that overexpression of S1PR1 reversed the negative migration and invasion effects of miR-363 mimics in 786O cells. f Knockdown of S1PR1 counteracted the positive migration and invasion effects of miR-363 inhibitor in 769P cells. g Alterations of S1PR1, ERK and downstream genes of ERK protein level in 786O cells transfected with miR-363 mimic (versus NC mimics) and lentiviral S1PR1 plasmids (versus empty vector). h Alterations of S1PR1, ERK and downstream genes of ERK protein level in 769P cells transfected with miR-363 inhibitor (versus NC inhibitor) and siS1PR1 (versus siNC). Data are presented as the mean ± SD. (*P

    Article Snippet: Then, the cells were incubated with primary antibody against S1PR1 (ab-11424; Abcam, Cambridge, UK) at a 1:50 dilution.

    Techniques: Migration, Transfection, Plasmid Preparation, MTS Assay, Over Expression

    Overexpression of miR-363 suppresses xenograft tumour growth in vivo. a Xenograft tumours were obtained and dissected 8 weeks after subcutaneous injection of 786O cells stably transfected with lentiviral miR-363 particles or empty vector. b Comparison of tumour volume and weight between the miR-363 overexpression group and the EV group (10 mice per group). c , d Alterations in the protein levels of S1PR1, ERK and downstream genes of ERK in xenograft tumours between the miR-363 overexpression group and the EV group. e Histoscores of S1PR1, t-ERK and p-ERK in IHC-stained xenograft tumours between the miR-363 overexpression group and the EV group. f Representative IHC staining images of S1PR1, t-ERK, p-ERK and negative control in xenograft tumours between the miR-363 overexpression group and EV group. Data are presented as the mean ± SD. (*P

    Journal: Cancer Cell International

    Article Title: miR-363 suppresses the proliferation, migration and invasion of clear cell renal cell carcinoma by downregulating S1PR1

    doi: 10.1186/s12935-020-01313-9

    Figure Lengend Snippet: Overexpression of miR-363 suppresses xenograft tumour growth in vivo. a Xenograft tumours were obtained and dissected 8 weeks after subcutaneous injection of 786O cells stably transfected with lentiviral miR-363 particles or empty vector. b Comparison of tumour volume and weight between the miR-363 overexpression group and the EV group (10 mice per group). c , d Alterations in the protein levels of S1PR1, ERK and downstream genes of ERK in xenograft tumours between the miR-363 overexpression group and the EV group. e Histoscores of S1PR1, t-ERK and p-ERK in IHC-stained xenograft tumours between the miR-363 overexpression group and the EV group. f Representative IHC staining images of S1PR1, t-ERK, p-ERK and negative control in xenograft tumours between the miR-363 overexpression group and EV group. Data are presented as the mean ± SD. (*P

    Article Snippet: Then, the cells were incubated with primary antibody against S1PR1 (ab-11424; Abcam, Cambridge, UK) at a 1:50 dilution.

    Techniques: Over Expression, In Vivo, Injection, Stable Transfection, Transfection, Plasmid Preparation, Mouse Assay, Immunohistochemistry, Staining, Negative Control

    Expression and prognostic significance of miR-363 in ccRCC and its relationship with S1PR1 expression. a miR-363 expression levels were significantly downregulated in ccRCC tissues compared to adjacent normal tissues. b–d Comparison of miR-363 expression levels between subgroups of patients by clinical stage, T stage and Fuhrman grade. e miR-363 expression in normal renal cell lines and various RCC cell lines. f Kaplan–Meier analysis of ccRCC patients in the low miR-363 group (n = 39) and the high miR-363 group (n = 38) with regard to disease-free survival. g S1PR1 expression levels were markedly upregulated in ccRCC tissues compared to adjacent normal tissues. h and i S1PR1 protein expression in normal renal cell lines and various RCC cell lines. j Negative correlation between S1PR1 mRNA levels and miR-363 levels in ccRCC tissues (n = 77, r = −0.509, P

    Journal: Cancer Cell International

    Article Title: miR-363 suppresses the proliferation, migration and invasion of clear cell renal cell carcinoma by downregulating S1PR1

    doi: 10.1186/s12935-020-01313-9

    Figure Lengend Snippet: Expression and prognostic significance of miR-363 in ccRCC and its relationship with S1PR1 expression. a miR-363 expression levels were significantly downregulated in ccRCC tissues compared to adjacent normal tissues. b–d Comparison of miR-363 expression levels between subgroups of patients by clinical stage, T stage and Fuhrman grade. e miR-363 expression in normal renal cell lines and various RCC cell lines. f Kaplan–Meier analysis of ccRCC patients in the low miR-363 group (n = 39) and the high miR-363 group (n = 38) with regard to disease-free survival. g S1PR1 expression levels were markedly upregulated in ccRCC tissues compared to adjacent normal tissues. h and i S1PR1 protein expression in normal renal cell lines and various RCC cell lines. j Negative correlation between S1PR1 mRNA levels and miR-363 levels in ccRCC tissues (n = 77, r = −0.509, P

    Article Snippet: Then, the cells were incubated with primary antibody against S1PR1 (ab-11424; Abcam, Cambridge, UK) at a 1:50 dilution.

    Techniques: Expressing

    miR-363 downregulates S1PR1 expression by directly targeting its 3′-UTR. a S1PR1 mRNA level changes in 786O and 769P cells treated with different interferences. b , c Overexpression of miR-363 decreased the expression of S1PR1 protein levels in ccRCC cells, whereas knockdown of miR-363 increased the expression of S1PR1 protein levels in ccRCC cells. d Representative immunofluorescent staining images showed the inverse effect of miR-363 on S1PR1 in ccRCC cells. e Sequence alignment of the S1PR1 3′-UTR with wild-type (WT) versus mutant (MUT) predicted potential miR-363 binding sites. f Luciferase reporter assay showed attenuated reporter activity after transfection of the wild-type S1PR1 3′-UTR reporter construct in human embryonic kidney 293T cells overexpressing miR-363. The S1PR1 3′-UTR MUT and control constructs had no effect on reporter activity. Data are presented as the mean ± SD. (*P

    Journal: Cancer Cell International

    Article Title: miR-363 suppresses the proliferation, migration and invasion of clear cell renal cell carcinoma by downregulating S1PR1

    doi: 10.1186/s12935-020-01313-9

    Figure Lengend Snippet: miR-363 downregulates S1PR1 expression by directly targeting its 3′-UTR. a S1PR1 mRNA level changes in 786O and 769P cells treated with different interferences. b , c Overexpression of miR-363 decreased the expression of S1PR1 protein levels in ccRCC cells, whereas knockdown of miR-363 increased the expression of S1PR1 protein levels in ccRCC cells. d Representative immunofluorescent staining images showed the inverse effect of miR-363 on S1PR1 in ccRCC cells. e Sequence alignment of the S1PR1 3′-UTR with wild-type (WT) versus mutant (MUT) predicted potential miR-363 binding sites. f Luciferase reporter assay showed attenuated reporter activity after transfection of the wild-type S1PR1 3′-UTR reporter construct in human embryonic kidney 293T cells overexpressing miR-363. The S1PR1 3′-UTR MUT and control constructs had no effect on reporter activity. Data are presented as the mean ± SD. (*P

    Article Snippet: Then, the cells were incubated with primary antibody against S1PR1 (ab-11424; Abcam, Cambridge, UK) at a 1:50 dilution.

    Techniques: Expressing, Over Expression, Staining, Sequencing, Mutagenesis, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Transfection, Construct

    S1PR1 promotes the proliferation, migration and invasion of ccRCC cells in vitro. a Folds change of S1PR1 expression levels in 786O cells after knockdown with siRNA. b Folds change of the S1PR1 expression levels in 769P cells after overexpression with lentiviral S1PR1 plasmids. c MTS assay suggested that knockdown of S1PR1 inhibited the proliferation of 786O cells and that overexpression of S1PR1 promoted the proliferation of 769P cells. d Colony formation assay suggested that knockdown of S1PR1 inhibited the colony number in 786O cells and that overexpression of S1PR1 promoted the colony number in 769P cells. e Transwell assays suggested that knockdown of S1PR1 inhibits migration and invasion in 786O cells and that overexpression of S1PR1 promoted migration and invasion in 769P cells. f Wound healing assay suggested that knockdown of S1PR1 inhibited the cell mobility of 786O cells and that overexpression of S1PR1 promoted the cell mobility of 769P cells. Data are presented as the mean ± SD. (*P

    Journal: Cancer Cell International

    Article Title: miR-363 suppresses the proliferation, migration and invasion of clear cell renal cell carcinoma by downregulating S1PR1

    doi: 10.1186/s12935-020-01313-9

    Figure Lengend Snippet: S1PR1 promotes the proliferation, migration and invasion of ccRCC cells in vitro. a Folds change of S1PR1 expression levels in 786O cells after knockdown with siRNA. b Folds change of the S1PR1 expression levels in 769P cells after overexpression with lentiviral S1PR1 plasmids. c MTS assay suggested that knockdown of S1PR1 inhibited the proliferation of 786O cells and that overexpression of S1PR1 promoted the proliferation of 769P cells. d Colony formation assay suggested that knockdown of S1PR1 inhibited the colony number in 786O cells and that overexpression of S1PR1 promoted the colony number in 769P cells. e Transwell assays suggested that knockdown of S1PR1 inhibits migration and invasion in 786O cells and that overexpression of S1PR1 promoted migration and invasion in 769P cells. f Wound healing assay suggested that knockdown of S1PR1 inhibited the cell mobility of 786O cells and that overexpression of S1PR1 promoted the cell mobility of 769P cells. Data are presented as the mean ± SD. (*P

    Article Snippet: Then, the cells were incubated with primary antibody against S1PR1 (ab-11424; Abcam, Cambridge, UK) at a 1:50 dilution.

    Techniques: Migration, In Vitro, Expressing, Over Expression, MTS Assay, Colony Assay, Wound Healing Assay

    S1P activates NF-κB via S1PR1 and S1PR2. (A and B) M2 and A7 melanoma cells were serum starved overnight. (A) RNA was isolated, and levels of S1PR1 to -3 mRNAs were quantified by QPCR and normalized to GAPDH levels. Data are expressed as relative

    Journal: Molecular and Cellular Biology

    Article Title: Filamin A Expression Negatively Regulates Sphingosine-1-Phosphate-Induced NF-κB Activation in Melanoma Cells by Inhibition of Akt Signaling

    doi: 10.1128/MCB.00554-15

    Figure Lengend Snippet: S1P activates NF-κB via S1PR1 and S1PR2. (A and B) M2 and A7 melanoma cells were serum starved overnight. (A) RNA was isolated, and levels of S1PR1 to -3 mRNAs were quantified by QPCR and normalized to GAPDH levels. Data are expressed as relative

    Article Snippet: S1PR1, S1PR2, and S1PR3 antibodies were obtained from Abcam (Cambridge, MA).

    Techniques: Isolation, Real-time Polymerase Chain Reaction