Review

rabbit polyclonal anti sphk1 (Proteintech)

Name: rabbit polyclonal anti sphk1
Brand: Proteintech
Rating: 95 (81 reviews)

Proteintech is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Proteintech rabbit polyclonal anti sphk1
Rabbit Polyclonal Anti Sphk1, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti sphk1/product/Proteintech
Average 95 stars, based on 81 article reviews

rabbit polyclonal anti sphk1 - by Bioz Stars, 2026-02

95/100 stars

Images

Image Search Results

Spatial transcriptome map of internal hemorrhoid tissue. A: Hematoxylin and eosin-stained section of internal hemorrhoid tissue; B: Spatial transcriptome; C: Differences in expression of the classic marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in fibroblasts; D: Spatial localization of the classic marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in fibroblasts; E: Spatial distribution of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1; F: Differences in gene expression of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1. COL3A1 : Collagen 3 alpha 1; DCN : Decorin; ACTA2 : Actin alpha 2; Sphk1 : Sphingosine kinase 1; S1PR1 : Sphingosine-1-phosphate receptor 1; TGF-β1 : Transforming growth factor-β1.

Journal: World Journal of Gastrointestinal Surgery

Article Title: Single-nucleus RNA sequencing and spatial transcriptomics reveal the mechanism by which Xiaozhiling injection treats internal hemorrhoids

doi: 10.4240/wjgs.v17.i4.103494

Figure Lengend Snippet: Spatial transcriptome map of internal hemorrhoid tissue. A: Hematoxylin and eosin-stained section of internal hemorrhoid tissue; B: Spatial transcriptome; C: Differences in expression of the classic marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in fibroblasts; D: Spatial localization of the classic marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in fibroblasts; E: Spatial distribution of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1; F: Differences in gene expression of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1. COL3A1 : Collagen 3 alpha 1; DCN : Decorin; ACTA2 : Actin alpha 2; Sphk1 : Sphingosine kinase 1; S1PR1 : Sphingosine-1-phosphate receptor 1; TGF-β1 : Transforming growth factor-β1.

Article Snippet: Xiaozhiling injection (Beijing China Resources High-Tech Natural Medicine Co., Ltd.); inhaled isoflurane (Baxter Healthcare Corporation, China); 4% neutral formaldehyde fixative solution; 6% croton oil (Beijing Huawei Ruike Chemical Co., Ltd.); pyridine (Beijing Lambolide Trading Co., Ltd.); ether (Xilong Science Co., Ltd.); horseradish peroxidase-conjugated polyclonal anti-rabbit/mouse IgG (Boster); an anti-TGF-β1 rabbit polyclonal antibody (Proteintech); an anti-Sphk1 rabbit polyclonal antibody (Proteintech); an anti-sphingosine-1-phosphate receptor 1 (S1PR1) rabbit polyclonal antibody (Proteintech); and an alpha smooth muscle actin specific monoclonal antibody (Proteintech) were used in this study.

Techniques: Staining, Expressing, Marker, Gene Expression

Spatial transcriptome. A: Hematoxylin and eosin-stained section 1 week after Xiaozhiling injection; B: Spatial transcriptome; C: Differences in expression of the classic fibroblast marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in different clusters; D: Spatial localization of the classic fibroblast marker genes collagen 3 alpha 1, decorin, and actin alpha 2; E: Spatial localization of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1; F: Differences in expression of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1 in different clusters; G: Analysis of differences between the two groups. COL3A1 : Collagen 3 alpha 1; DCN : Decorin; ACTA2 : Actin alpha 2; Sphk1: Sphingosine kinase 1; S1PR1: Sphingosine-1-phosphate receptor 1; TGF-β1: Transforming growth factor-β1.

Journal: World Journal of Gastrointestinal Surgery

Article Title: Single-nucleus RNA sequencing and spatial transcriptomics reveal the mechanism by which Xiaozhiling injection treats internal hemorrhoids

doi: 10.4240/wjgs.v17.i4.103494

Figure Lengend Snippet: Spatial transcriptome. A: Hematoxylin and eosin-stained section 1 week after Xiaozhiling injection; B: Spatial transcriptome; C: Differences in expression of the classic fibroblast marker genes collagen 3 alpha 1, decorin, and actin alpha 2 in different clusters; D: Spatial localization of the classic fibroblast marker genes collagen 3 alpha 1, decorin, and actin alpha 2; E: Spatial localization of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1; F: Differences in expression of sphingosine kinase 1, sphingosine-1-phosphate receptor 1, and transforming growth factor-β1 in different clusters; G: Analysis of differences between the two groups. COL3A1 : Collagen 3 alpha 1; DCN : Decorin; ACTA2 : Actin alpha 2; Sphk1: Sphingosine kinase 1; S1PR1: Sphingosine-1-phosphate receptor 1; TGF-β1: Transforming growth factor-β1.

Techniques: Staining, Injection, Expressing, Marker

Immunohistochemical staining of internal hemorrhoid tissue. A: Sphingosine kinase 1 (100 ×); B: Sphingosine-1-phosphate receptor 1 (100 ×); C: Transforming growth factor-β1 (100 ×); D: Sphingosine kinase 1 (400 ×); E: Sphingosine-1-phosphate receptor 1 (400 ×); F: Transforming growth factor-β1 (400 ×).

Journal: World Journal of Gastrointestinal Surgery

Article Title: Single-nucleus RNA sequencing and spatial transcriptomics reveal the mechanism by which Xiaozhiling injection treats internal hemorrhoids

doi: 10.4240/wjgs.v17.i4.103494

Figure Lengend Snippet: Immunohistochemical staining of internal hemorrhoid tissue. A: Sphingosine kinase 1 (100 ×); B: Sphingosine-1-phosphate receptor 1 (100 ×); C: Transforming growth factor-β1 (100 ×); D: Sphingosine kinase 1 (400 ×); E: Sphingosine-1-phosphate receptor 1 (400 ×); F: Transforming growth factor-β1 (400 ×).

Techniques: Immunohistochemical staining, Staining

Immunohistochemical staining. A: Sphingosine kinase 1 (100 ×); B: Sphingosine-1-phosphate receptor 1 (100 ×); C: Transforming growth factor-β1 (100 ×); D: Sphingosine kinase 1 (400 ×); E: Sphingosine-1-phosphate receptor 1 (400 ×); F: Transforming growth factor-β1 (400 ×).

Journal: World Journal of Gastrointestinal Surgery

Article Title: Single-nucleus RNA sequencing and spatial transcriptomics reveal the mechanism by which Xiaozhiling injection treats internal hemorrhoids

doi: 10.4240/wjgs.v17.i4.103494

Figure Lengend Snippet: Immunohistochemical staining. A: Sphingosine kinase 1 (100 ×); B: Sphingosine-1-phosphate receptor 1 (100 ×); C: Transforming growth factor-β1 (100 ×); D: Sphingosine kinase 1 (400 ×); E: Sphingosine-1-phosphate receptor 1 (400 ×); F: Transforming growth factor-β1 (400 ×).

Techniques: Immunohistochemical staining, Staining

Expression of sphingosine kinase 1-sphingosine-1-phosphate pathway related proteins in rectal tissues. Sphk1: Sphingosine kinase 1; S1PR1: Sphingosine-1-phosphate receptor 1; TGF-β1: Transforming growth factor-β1.

Journal: World Journal of Gastrointestinal Surgery

Article Title: Single-nucleus RNA sequencing and spatial transcriptomics reveal the mechanism by which Xiaozhiling injection treats internal hemorrhoids

doi: 10.4240/wjgs.v17.i4.103494

Figure Lengend Snippet: Expression of sphingosine kinase 1-sphingosine-1-phosphate pathway related proteins in rectal tissues. Sphk1: Sphingosine kinase 1; S1PR1: Sphingosine-1-phosphate receptor 1; TGF-β1: Transforming growth factor-β1.

Techniques: Expressing

The production of sphingosine 1-phosphate (S1P) and ceramide 1-phosphate (C1P) is vital for sperm capacitation. Effect of PF453 (SphK1 inhibitor), NVP213 (CERK inhibitor), and SLM (SphK2 inhibitor) on tyrosine phosphorylation (P-Tyr) ( a and b ) and phospho-SphK1 (P-SphK1) fluorescence ( c ). Spermatozoa were incubated for 3.5 h under capacitating conditions without any treatment (control) or the presence of FCSu (10% v/v), with or without PF453, NVP213, and SLM. (a) The immunoblotting analysis shows decreased levels of P-Tyr in human spermatozoa treated with increasing concentrations (100 nM and 1 μM) of SphK1 and CERK inhibitors. (b) Inhibition of SphK2 with SLM did not change the levels of P-Tyr. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001).

Journal: Human Reproduction (Oxford, England)

Article Title: Sphingolipids modulate redox signalling during human sperm capacitation

doi: 10.1093/humrep/deae268

Figure Lengend Snippet: The production of sphingosine 1-phosphate (S1P) and ceramide 1-phosphate (C1P) is vital for sperm capacitation. Effect of PF453 (SphK1 inhibitor), NVP213 (CERK inhibitor), and SLM (SphK2 inhibitor) on tyrosine phosphorylation (P-Tyr) ( a and b ) and phospho-SphK1 (P-SphK1) fluorescence ( c ). Spermatozoa were incubated for 3.5 h under capacitating conditions without any treatment (control) or the presence of FCSu (10% v/v), with or without PF453, NVP213, and SLM. (a) The immunoblotting analysis shows decreased levels of P-Tyr in human spermatozoa treated with increasing concentrations (100 nM and 1 μM) of SphK1 and CERK inhibitors. (b) Inhibition of SphK2 with SLM did not change the levels of P-Tyr. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001).

Article Snippet: Rabbit polyclonal anti-P-SphK1 antibody was purchased from Invitrogen (Fisher Scientific, Ottawa, ON, Canada).

Techniques: Fluorescence, Incubation, Control, Western Blot, Inhibition

Ceramide 1-phosphate (C1P) is required for the activation of SphK1 and sphingosine 1-phosphate (S1P) production. ( a ) Immunocytochemistry analysis of permeabilized spermatozoa showed the increase in fluorescence intensity of P-SphK1 with FCSu and diminished signal following inhibition with NVP231 (scale bar = 20 μM). ( b ) For the mean fluorescence intensity in arbitrary unit (a.u.), 200 cells were evaluated in the permeabilized samples. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; ** P ≤ 0.01).

Journal: Human Reproduction (Oxford, England)

Article Title: Sphingolipids modulate redox signalling during human sperm capacitation

doi: 10.1093/humrep/deae268

Figure Lengend Snippet: Ceramide 1-phosphate (C1P) is required for the activation of SphK1 and sphingosine 1-phosphate (S1P) production. ( a ) Immunocytochemistry analysis of permeabilized spermatozoa showed the increase in fluorescence intensity of P-SphK1 with FCSu and diminished signal following inhibition with NVP231 (scale bar = 20 μM). ( b ) For the mean fluorescence intensity in arbitrary unit (a.u.), 200 cells were evaluated in the permeabilized samples. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; ** P ≤ 0.01).

Article Snippet: Rabbit polyclonal anti-P-SphK1 antibody was purchased from Invitrogen (Fisher Scientific, Ottawa, ON, Canada).

Techniques: Activation Assay, Immunocytochemistry, Fluorescence, Inhibition

Sphingolipids stimulate NO • production during capacitation. Foetal cord serum ultrafiltrate (FCSu)-, sphingosine (Sph)-, and ceramide (Cer)-capacitated spermatozoa incubated with or without L-NAME (nitric oxide synthase ‘NOS’ inhibitor), NVP231 (CERK inhibitor), PF543 (SphK1 inhibitor), and VCP23019 (S1PR1 inhibitor), on tyrosine phosphorylation (P-Tyr) and DAF-2DA (probe for intracellular nitric oxide (NO • )) and Sytox Blue (viability dye) fluorescence. ( a ) The immunoblotting analysis shows a decrease in P-Tyr with L-NAME. Each lane was normalized to its silver-stain optical density value for signal quantification. ( b ) Histogram of viable spermatozoa (Sytox Blue negative) that are either DAF2-DA + (blue, P1) or DAF2-DA − (yellow, P3) for NO • production. These spermatozoa were incubated with the inducers FCSu, Sph, and Cer, and were treated with VCP23019, NVP231, and PF543. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; *** P ≤ 0.001).

Journal: Human Reproduction (Oxford, England)

Article Title: Sphingolipids modulate redox signalling during human sperm capacitation

doi: 10.1093/humrep/deae268

Figure Lengend Snippet: Sphingolipids stimulate NO • production during capacitation. Foetal cord serum ultrafiltrate (FCSu)-, sphingosine (Sph)-, and ceramide (Cer)-capacitated spermatozoa incubated with or without L-NAME (nitric oxide synthase ‘NOS’ inhibitor), NVP231 (CERK inhibitor), PF543 (SphK1 inhibitor), and VCP23019 (S1PR1 inhibitor), on tyrosine phosphorylation (P-Tyr) and DAF-2DA (probe for intracellular nitric oxide (NO • )) and Sytox Blue (viability dye) fluorescence. ( a ) The immunoblotting analysis shows a decrease in P-Tyr with L-NAME. Each lane was normalized to its silver-stain optical density value for signal quantification. ( b ) Histogram of viable spermatozoa (Sytox Blue negative) that are either DAF2-DA + (blue, P1) or DAF2-DA − (yellow, P3) for NO • production. These spermatozoa were incubated with the inducers FCSu, Sph, and Cer, and were treated with VCP23019, NVP231, and PF543. The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; *** P ≤ 0.001).

Article Snippet: Rabbit polyclonal anti-P-SphK1 antibody was purchased from Invitrogen (Fisher Scientific, Ottawa, ON, Canada).

Techniques: Incubation, Fluorescence, Western Blot, Silver Staining

PKR activates SphK1 during human sperm capacitation. Foetal cord serum ultrafiltrate (FCSu)-, sphingosine (Sph)-, and ceramide (Cer)-capacitated spermatozoa incubated with or without protein kinase R (PKR) inhibitor (PKRI) were assessed for their impact on both tyrosine phosphorylation (P-Tyr), SphK1 phosphorylation (P-SphK1), and PKR phosphorylation (P-PKR) fluorescence. ( a ) The immunoblotting analysis demonstrates decreased P-SphK1 and corresponding P-Tyr levels in capacitated samples treated with PKRI. ( b and c ) Immunocytochemistry analysis shows an increase in P-PKR activation upon induction of capacitation as measured by mean fluorescence (a.u.). For immunocytochemistry analysis, 200 cells were analysed per permeabilized sperm sample (scale bar = 5 μM). The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001).

Journal: Human Reproduction (Oxford, England)

Article Title: Sphingolipids modulate redox signalling during human sperm capacitation

doi: 10.1093/humrep/deae268

Figure Lengend Snippet: PKR activates SphK1 during human sperm capacitation. Foetal cord serum ultrafiltrate (FCSu)-, sphingosine (Sph)-, and ceramide (Cer)-capacitated spermatozoa incubated with or without protein kinase R (PKR) inhibitor (PKRI) were assessed for their impact on both tyrosine phosphorylation (P-Tyr), SphK1 phosphorylation (P-SphK1), and PKR phosphorylation (P-PKR) fluorescence. ( a ) The immunoblotting analysis demonstrates decreased P-SphK1 and corresponding P-Tyr levels in capacitated samples treated with PKRI. ( b and c ) Immunocytochemistry analysis shows an increase in P-PKR activation upon induction of capacitation as measured by mean fluorescence (a.u.). For immunocytochemistry analysis, 200 cells were analysed per permeabilized sperm sample (scale bar = 5 μM). The results represent sperm samples from different healthy donors (n = 4, ANOVA and Tukey test; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001).

Article Snippet: Rabbit polyclonal anti-P-SphK1 antibody was purchased from Invitrogen (Fisher Scientific, Ottawa, ON, Canada).

Techniques: Incubation, Fluorescence, Western Blot, Immunocytochemistry, Activation Assay

Schematic of the sphingolipid signalling cascade during capacitation. Following the production of the bioactive sphingolipid, ceramide (Cer), through the action of a neutral sphingomyelinase (nSMase), Cer will engage with both the ceramidase (CDase) and ceramide kinase (CERK), yielding sphingosine (Sph), and ceramide 1-phosphate (C1P), respectively. Sphingosine (sph) and Cer will lead to a spike in superoxide ( O 2 •− ) production by the sperm oxidase and protein kinase R (PKR) will auto-phosphorylate (P-PKR). C1P, PKC, and P-PKR will facilitate the translocation of SphK1 to the inner leaflet of the plasma membrane and activation by phosphorylation (Ser 225 ). P-SphK1 will convert Sph to sphingosine 1-phosphate (S1P), and the latter will interact with the ABCC1 transporter and efflux to the outer leaflet. S1P will interact with the S1PR1 and initiate the G i signalling cascade. Signal transduction activates the PI3K-AKT and phospholipase C (PLC)-PKC pathways that lead to the subsequent activation of nitric oxide synthase (NOS) and nitric oxide (NO • ) production and SphK1, respectively. NO • interacts with Ras to facilitate the extracellular signal-regulated kinase (ERK) signalling cascade leading to an increase in tyrosine phosphorylation (P-Tyr) of prominent sperm proteins required for capacitation. The figure was created with BioRender.com.

Journal: Human Reproduction (Oxford, England)

Article Title: Sphingolipids modulate redox signalling during human sperm capacitation

doi: 10.1093/humrep/deae268

Figure Lengend Snippet: Schematic of the sphingolipid signalling cascade during capacitation. Following the production of the bioactive sphingolipid, ceramide (Cer), through the action of a neutral sphingomyelinase (nSMase), Cer will engage with both the ceramidase (CDase) and ceramide kinase (CERK), yielding sphingosine (Sph), and ceramide 1-phosphate (C1P), respectively. Sphingosine (sph) and Cer will lead to a spike in superoxide ( O 2 •− ) production by the sperm oxidase and protein kinase R (PKR) will auto-phosphorylate (P-PKR). C1P, PKC, and P-PKR will facilitate the translocation of SphK1 to the inner leaflet of the plasma membrane and activation by phosphorylation (Ser 225 ). P-SphK1 will convert Sph to sphingosine 1-phosphate (S1P), and the latter will interact with the ABCC1 transporter and efflux to the outer leaflet. S1P will interact with the S1PR1 and initiate the G i signalling cascade. Signal transduction activates the PI3K-AKT and phospholipase C (PLC)-PKC pathways that lead to the subsequent activation of nitric oxide synthase (NOS) and nitric oxide (NO • ) production and SphK1, respectively. NO • interacts with Ras to facilitate the extracellular signal-regulated kinase (ERK) signalling cascade leading to an increase in tyrosine phosphorylation (P-Tyr) of prominent sperm proteins required for capacitation. The figure was created with BioRender.com.

Article Snippet: Rabbit polyclonal anti-P-SphK1 antibody was purchased from Invitrogen (Fisher Scientific, Ottawa, ON, Canada).

Techniques: Translocation Assay, Membrane, Activation Assay, Transduction

Journal: Journal of Inflammation Research

Article Title: Rosmarinic Acid Alleviates Radiation-Induced Pulmonary Fibrosis by Downregulating the tRNA N7-Methylguanosine Modification-Regulated Fibroblast-to-Myofibroblast Transition Through the Exosome Pathway

doi: 10.2147/JIR.S458794

Figure Lengend Snippet: RA reversed the increase in tRNA expression and m7G modification levels in irradiated lung epithelial cells. ( A ) Quantification of the m7G modification level of tRNAs. ( B ) Expression profile of tRNAs. Fold Change cut-off: 1.5, P-value cut-off: 0.05. ( C and D ) Northwestern and northern blotting of the indicated tRNAs was performed in TC-1 cells and mouse lung tissue. ( E ) The expression levels of the METTL1/WDR4 complex were evaluated via Western blotting. ( F ) The efficiency of both knockdown and overexpression of METTL1 and WDR4 genes were confirmed by qRT‒PCR. ( G ) The effects of METTL1 and WDR4 on tRNA expression and m7G modification were detected via northwestern and northern blotting. The precise n value (number of biologically independent replicates) is 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The following antibodies were used: METTL1 polyclonal antibody (Proteintech, 14994-1-AP), rabbit anti-WDR4 polyclonal antibody (Absin, abs152662), SPHK1 polyclonal antibody (Proteintech, 10670-1-AP), N-acetyltransferase 10 (NAT10) polyclonal antibody (Proteintech; 13365-1-AP), acetyl-lysine (Affinity, DF7729), PFKFB3 antibody (Affinity, DF12016), phospho-PFKFB3 (Ser461) antibody (Affinity, AF3581), phospho-AMPK alpha (Thr172) antibody (Affinity, AF3423), AMPK alpha antibody (Affinity, AF6423), and ACTIN antibody (Proteintech, 81115-1-RR).

Techniques: Expressing, Modification, Irradiation, Northern Blot, Western Blot, Knockdown, Over Expression

RA affected the METTL1/WDR4-mediated tRNA m7G modification of MLFs via exosomes. ( A and B ) Expression levels of representative m7G-modified tRNAs in exosomes were examined by northwestern and northern blotting using the indicated tRNA probes. ( C ) MLFs were incubated with exosomes derived from TC-1 cells, and the expression and m7G modification levels of the indicated tRNAs were detected via northwestern and northern blot assays. The precise n value (number of biologically independent replicates) is 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: Journal of Inflammation Research

doi: 10.2147/JIR.S458794

Figure Lengend Snippet: RA affected the METTL1/WDR4-mediated tRNA m7G modification of MLFs via exosomes. ( A and B ) Expression levels of representative m7G-modified tRNAs in exosomes were examined by northwestern and northern blotting using the indicated tRNA probes. ( C ) MLFs were incubated with exosomes derived from TC-1 cells, and the expression and m7G modification levels of the indicated tRNAs were detected via northwestern and northern blot assays. The precise n value (number of biologically independent replicates) is 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Techniques: Modification, Expressing, Northern Blot, Incubation, Derivative Assay

RA suppressed the process of FMT in MLFs by downregulating METTL1/WDR4-mediated tRNA m7G modification via exosomes of TC-1 cells. ( A ) The expression of α-SMA was assessed via immunofluorescence staining. Scale bars, 50 μm. ( B ) Cell viability was tested via EdU staining. Scale bars, 50 μm. ( C ) Migration ability was detected by a wound healing assay. Scale bars, 50 μm. The precise n value (number of biologically independent replicates) is 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: Journal of Inflammation Research

doi: 10.2147/JIR.S458794

Figure Lengend Snippet: RA suppressed the process of FMT in MLFs by downregulating METTL1/WDR4-mediated tRNA m7G modification via exosomes of TC-1 cells. ( A ) The expression of α-SMA was assessed via immunofluorescence staining. Scale bars, 50 μm. ( B ) Cell viability was tested via EdU staining. Scale bars, 50 μm. ( C ) Migration ability was detected by a wound healing assay. Scale bars, 50 μm. The precise n value (number of biologically independent replicates) is 3. *P < 0.05, **P < 0.01, ***P < 0.001.

Techniques: Modification, Expressing, Immunofluorescence, Staining, Migration, Wound Healing Assay

(A) Schematic representation of the regulation of S1P homeostasis by sphingosine kinase 1/2 (SphK1/2), S1P phosphatase 1/2 (SGPP1/2), S1P lyase (SGPL1) and S1P transporter Spinster 2 (Spns2). (B-G) Fold change in expression of SPHK1 (B), SPHK2 (C), SGPL1 (D), SGPP1 (E), SGPP2 (F) and SPNS2 (G) in hCMEC/D3s infected with N . meningitidis MC58 for an 8h infection time course normalized to 18S rRNA measured by qPCR. Data represent mean ± SD of 3 independent experiments measured in duplicates. Comparison to control for each time point using multiple t-test and p-value adjustment with Holm-Sidak correction. ***p<0.001 vs. respective control. (H) SphK enzymatic activity measured in hCMEC/D3s infected with N . meningitidis MC58 for a 6 h infection time in comparison to uninfected control cells as determined by ATP depletion assay. Uninfected control cells without addition of SphK substrate Sph (‘-Sph’) were used to detect non-specific ATP consumption. Graph represents the mean ± SD of at least three independent experiments with biological duplicates and is reported as activity normalized to control cells. One-Way ANOVA followed by by Dunett’s post-hoc test. *p<0.05, **p<0.01, ****p<0.0001 vs. control. (I) hCMEC/D3s were infected with N . meningitidis MC58 for 8h infection time course and both total protein and/or phosphorylated form of SphK1 (pSer-225) and SphK2 (pThr-578) were assessed by Western blot. Representative immunoblot is shown. Graphs represent the mean ± SD of three independent experiments and reported as phosphorylated form relative to total target protein normalized to control cells. One-Way ANOVA showed no significance. Related to .

Journal: PLOS Pathogens

Article Title: Sphingosine kinase 1/S1P receptor signaling axis is essential for cellular uptake of Neisseria meningitidis in brain endothelial cells

doi: 10.1371/journal.ppat.1011842

Figure Lengend Snippet: (A) Schematic representation of the regulation of S1P homeostasis by sphingosine kinase 1/2 (SphK1/2), S1P phosphatase 1/2 (SGPP1/2), S1P lyase (SGPL1) and S1P transporter Spinster 2 (Spns2). (B-G) Fold change in expression of SPHK1 (B), SPHK2 (C), SGPL1 (D), SGPP1 (E), SGPP2 (F) and SPNS2 (G) in hCMEC/D3s infected with N . meningitidis MC58 for an 8h infection time course normalized to 18S rRNA measured by qPCR. Data represent mean ± SD of 3 independent experiments measured in duplicates. Comparison to control for each time point using multiple t-test and p-value adjustment with Holm-Sidak correction. ***p<0.001 vs. respective control. (H) SphK enzymatic activity measured in hCMEC/D3s infected with N . meningitidis MC58 for a 6 h infection time in comparison to uninfected control cells as determined by ATP depletion assay. Uninfected control cells without addition of SphK substrate Sph (‘-Sph’) were used to detect non-specific ATP consumption. Graph represents the mean ± SD of at least three independent experiments with biological duplicates and is reported as activity normalized to control cells. One-Way ANOVA followed by by Dunett’s post-hoc test. *p<0.05, **p<0.01, ****p<0.0001 vs. control. (I) hCMEC/D3s were infected with N . meningitidis MC58 for 8h infection time course and both total protein and/or phosphorylated form of SphK1 (pSer-225) and SphK2 (pThr-578) were assessed by Western blot. Representative immunoblot is shown. Graphs represent the mean ± SD of three independent experiments and reported as phosphorylated form relative to total target protein normalized to control cells. One-Way ANOVA showed no significance. Related to .

Article Snippet: Rabbit polyclonal pospho-specific Anti-(Ser-225)-SphK1(ECM biosciences # SP1641) , 1:250 , 5% BSA in TBS-T.

Techniques: Expressing, Infection, Comparison, Activity Assay, ATP Depletion Assay, Western Blot

(A) hCMEC/D3 were co-transfected with SPHK1-siRNA (100 nM) and SPHK2-siRNA (25 nM) or transfected with scrambled siRNA (50 nM) for 24h and infected with N . meningitidis MC58 for 8h. Numbers of viable intracellular bacteria were determined by gentamicin protection assay and normalized to control cells (scrambled siRNA). Data are means ± SD; n = 4, N = 2. Unpaired students t-test, **** p < 0.0001. Absence of cytotoxicity and knockdown efficiency were verified as shown in . (B-D) hCMEC/D3 were treated with SphK1 inhibitor PF543 (B), SphK2 inhibitors SLM6031434 (C) or SphK2 inhibitor K145 (D) for 30 min or solvent control (DMSO) and subsequently infected with N . meningitidis MC58 for 8h. Numbers of viable intracellular bacteria were determined by gentamicin protection assay and normalized to control cells (DMSO treated cells). Data are means ± SD; n ≥ 4, N = 2. Comparison to DMSO-treated control using unpaired students t-test (A), multiple t-test with p-value adjustment with Holm-Sidak correction in comparison to control for each time point (B-D). ** p < 0.01, *** p < 0.001, **** p < 0.0001. Absence of cytotoxicity and bacteriostatic effects were verified as shown in .

Journal: PLOS Pathogens

Article Title: Sphingosine kinase 1/S1P receptor signaling axis is essential for cellular uptake of Neisseria meningitidis in brain endothelial cells

doi: 10.1371/journal.ppat.1011842

Figure Lengend Snippet: (A) hCMEC/D3 were co-transfected with SPHK1-siRNA (100 nM) and SPHK2-siRNA (25 nM) or transfected with scrambled siRNA (50 nM) for 24h and infected with N . meningitidis MC58 for 8h. Numbers of viable intracellular bacteria were determined by gentamicin protection assay and normalized to control cells (scrambled siRNA). Data are means ± SD; n = 4, N = 2. Unpaired students t-test, **** p < 0.0001. Absence of cytotoxicity and knockdown efficiency were verified as shown in . (B-D) hCMEC/D3 were treated with SphK1 inhibitor PF543 (B), SphK2 inhibitors SLM6031434 (C) or SphK2 inhibitor K145 (D) for 30 min or solvent control (DMSO) and subsequently infected with N . meningitidis MC58 for 8h. Numbers of viable intracellular bacteria were determined by gentamicin protection assay and normalized to control cells (DMSO treated cells). Data are means ± SD; n ≥ 4, N = 2. Comparison to DMSO-treated control using unpaired students t-test (A), multiple t-test with p-value adjustment with Holm-Sidak correction in comparison to control for each time point (B-D). ** p < 0.01, *** p < 0.001, **** p < 0.0001. Absence of cytotoxicity and bacteriostatic effects were verified as shown in .

Article Snippet: Rabbit polyclonal pospho-specific Anti-(Ser-225)-SphK1(ECM biosciences # SP1641) , 1:250 , 5% BSA in TBS-T.

Techniques: Transfection, Infection, Bacteria, Solvent, Comparison

Journal: PLOS Pathogens

Article Title: Sphingosine kinase 1/S1P receptor signaling axis is essential for cellular uptake of Neisseria meningitidis in brain endothelial cells

doi: 10.1371/journal.ppat.1011842

Figure Lengend Snippet: Primers used for qPCR.

Article Snippet: Rabbit polyclonal pospho-specific Anti-(Ser-225)-SphK1(ECM biosciences # SP1641) , 1:250 , 5% BSA in TBS-T.

Techniques: Sequencing

Journal: PLOS Pathogens

Article Title: Sphingosine kinase 1/S1P receptor signaling axis is essential for cellular uptake of Neisseria meningitidis in brain endothelial cells

doi: 10.1371/journal.ppat.1011842

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Rabbit polyclonal pospho-specific Anti-(Ser-225)-SphK1(ECM biosciences # SP1641) , 1:250 , 5% BSA in TBS-T.

Techniques: Milk

Expression of S1PR and SPK mRNA in multiple myeloma cell lines, HUVECs and primary samples. (A) SPK1 and SPK2 mRNA expression, and (B) the expression of five isotype S1P receptors (S1PR1-S1PR5) were assessed. HUVECs, human umbilical vessel cells; PCL, plasma cell leukemia; S1PR, sphingosine 1-phosphate receptor; SK, sphingosine kinase.

Journal: Oncology Letters

Article Title: Potential of a sphingosine 1-phosphate receptor antagonist and sphingosine kinase inhibitors as targets for multiple myeloma treatment

doi: 10.3892/ol.2022.13231

Figure Lengend Snippet: Expression of S1PR and SPK mRNA in multiple myeloma cell lines, HUVECs and primary samples. (A) SPK1 and SPK2 mRNA expression, and (B) the expression of five isotype S1P receptors (S1PR1-S1PR5) were assessed. HUVECs, human umbilical vessel cells; PCL, plasma cell leukemia; S1PR, sphingosine 1-phosphate receptor; SK, sphingosine kinase.

Article Snippet: Anti-S1PR1 and SphK1 rabbit polyclonal antibodies were purchased from Proteintech ® .

Techniques: Expressing, Clinical Proteomics

Effects of anti-sphingosine 1-phosphate agents on the growth of MM cell lines. (A) Fingolimod, SKI–I and ABC294640 suppressed the growth of the MM cell line (RPMI8226). (B) immunoblotting analysis was performed to measure the total protein extract after 24 h. *P<0.05 vs. control; # P<0.05 vs. 200 nM; ^ P<0.05 vs. 500 nM; **P<0.05 vs. 1 µM and ## P<0.05 vs. 10 µM. MM, multiple myeloma; PARP, poly (ADP-ribose) polymerase; SK, sphingosine kinase.

Journal: Oncology Letters

Article Title: Potential of a sphingosine 1-phosphate receptor antagonist and sphingosine kinase inhibitors as targets for multiple myeloma treatment

doi: 10.3892/ol.2022.13231

Figure Lengend Snippet: Effects of anti-sphingosine 1-phosphate agents on the growth of MM cell lines. (A) Fingolimod, SKI–I and ABC294640 suppressed the growth of the MM cell line (RPMI8226). (B) immunoblotting analysis was performed to measure the total protein extract after 24 h. *P<0.05 vs. control; # P<0.05 vs. 200 nM; ^ P<0.05 vs. 500 nM; **P<0.05 vs. 1 µM and ## P<0.05 vs. 10 µM. MM, multiple myeloma; PARP, poly (ADP-ribose) polymerase; SK, sphingosine kinase.

Article Snippet: Anti-S1PR1 and SphK1 rabbit polyclonal antibodies were purchased from Proteintech ® .

Techniques: Western Blot, Control

Cellular response of MM cell lines and HUVECs treated with carfilzomib, S1P and anti-S1P agents. (A) Effect of exogenous S1P on RPMI8226 cell growth inhibition by carfilzomib. (B) Effect of S1P addition during treatment with carfilzomib and the indicated inhibitor in different MM cell lines. *P<0.05. (C) Migration assay of HUVECs in the presence of a supernatant derived from a MM cell line. *P<0.05 cell line supernant vs. control. (D) Migration assay assessing the effect of S1P inhibitors on the S1P-mediated migration of HUVECs. *P<0.05 vs. S1P1 1μM. (E) Immunoblot assay showing the expression kinetic of pMAPK and ERK1 in HUVECs upon S1P addition. (F) Immunoblot assay showing the effect of S1P inhibitors on the expression of pMAPK. ABC, ABC294640; car, carfilzomib; Figo, fingolimod; HUVECs, human umbilical vessel cells; MM, multiple myeloma; p-, phosphorylated; S1P, sphingosine 1-phosphate; SK, sphingosine kinase.

Journal: Oncology Letters

Article Title: Potential of a sphingosine 1-phosphate receptor antagonist and sphingosine kinase inhibitors as targets for multiple myeloma treatment

doi: 10.3892/ol.2022.13231

Figure Lengend Snippet: Cellular response of MM cell lines and HUVECs treated with carfilzomib, S1P and anti-S1P agents. (A) Effect of exogenous S1P on RPMI8226 cell growth inhibition by carfilzomib. (B) Effect of S1P addition during treatment with carfilzomib and the indicated inhibitor in different MM cell lines. *P<0.05. (C) Migration assay of HUVECs in the presence of a supernatant derived from a MM cell line. *P<0.05 cell line supernant vs. control. (D) Migration assay assessing the effect of S1P inhibitors on the S1P-mediated migration of HUVECs. *P<0.05 vs. S1P1 1μM. (E) Immunoblot assay showing the expression kinetic of pMAPK and ERK1 in HUVECs upon S1P addition. (F) Immunoblot assay showing the effect of S1P inhibitors on the expression of pMAPK. ABC, ABC294640; car, carfilzomib; Figo, fingolimod; HUVECs, human umbilical vessel cells; MM, multiple myeloma; p-, phosphorylated; S1P, sphingosine 1-phosphate; SK, sphingosine kinase.

Article Snippet: Anti-S1PR1 and SphK1 rabbit polyclonal antibodies were purchased from Proteintech ® .

Techniques: Inhibition, Migration, Derivative Assay, Control, Western Blot, Expressing

Frequency of SphK1 and SphK2 IRS score. The frequency of immunoreactivity score (IRS) of sphingosine kinase 1 (SphK1) and sphingosine kinase 2 (SphK2) has been calculated with different clinicopathological attributes of oral squamous cell carcinoma patients. Tumor-node-metastasis (TNM) is grouped into two (Group I and Group II), whereas histology has been divided as moderately differentiated squamous cell carcinoma (MDSCC), well-differentiated squamous cell carcinoma (WDSCC), and poorly differentiated squamous cell carcinoma (PDSCC).

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: Frequency of SphK1 and SphK2 IRS score. The frequency of immunoreactivity score (IRS) of sphingosine kinase 1 (SphK1) and sphingosine kinase 2 (SphK2) has been calculated with different clinicopathological attributes of oral squamous cell carcinoma patients. Tumor-node-metastasis (TNM) is grouped into two (Group I and Group II), whereas histology has been divided as moderately differentiated squamous cell carcinoma (MDSCC), well-differentiated squamous cell carcinoma (WDSCC), and poorly differentiated squamous cell carcinoma (PDSCC).

Article Snippet: Rabbit polyclonal antibodies against human SphK1 (Cell Signaling Technology, Danvers, MA), human SphK2 (Cat# SAB1300017) (Sigma-Aldrich), human SGPP1 (Biorbyt, Cambridge, UK), and human LPP3 (Cat# HPA028892, Sigma-Aldrich) were used as the primary antibodies.

Techniques: Expressing

Western blotting for LPP3 (A-B), SphK2 (C), and SGPP1 (D) was performed in tumor tissues (T1, T2, T3, and T4) and adjacent normal tissues (N1, N2, N3, and N4) from the same patients. (E) Immunoblotting was performed in two sets of samples (N1, N2, T1, and T2). Photographs of the protein ladder were taken using Chemidoc under white light and were aligned with the immunoblots. Antibodies detected a single prominent band for each target protein, except LPP3, where two additional bands were observed on higher exposure (B). LPP3: lipid phosphate phosphatase 3; SphK2: sphingosine kinase 2; SGPP1: sphingosine-1-phosphate phosphatase 1; SphK1: sphingosine kinase 1; OSCC: oral squamous cell carcinoma.

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: Western blotting for LPP3 (A-B), SphK2 (C), and SGPP1 (D) was performed in tumor tissues (T1, T2, T3, and T4) and adjacent normal tissues (N1, N2, N3, and N4) from the same patients. (E) Immunoblotting was performed in two sets of samples (N1, N2, T1, and T2). Photographs of the protein ladder were taken using Chemidoc under white light and were aligned with the immunoblots. Antibodies detected a single prominent band for each target protein, except LPP3, where two additional bands were observed on higher exposure (B). LPP3: lipid phosphate phosphatase 3; SphK2: sphingosine kinase 2; SGPP1: sphingosine-1-phosphate phosphatase 1; SphK1: sphingosine kinase 1; OSCC: oral squamous cell carcinoma.

Techniques: Western Blot

(A-B) Neoplastic epithelium, (C) keratin pearl, (D) neural bundle, (E) skeletal muscle, and (F) blood vessels. N is non-neoplastic mucosa. SphK1: sphingosine kinase 1; OSCC: oral squamous cell carcinoma; IHC: immunohistochemistry.

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: (A-B) Neoplastic epithelium, (C) keratin pearl, (D) neural bundle, (E) skeletal muscle, and (F) blood vessels. N is non-neoplastic mucosa. SphK1: sphingosine kinase 1; OSCC: oral squamous cell carcinoma; IHC: immunohistochemistry.

Techniques: Immunohistochemistry

Expression of SphK2 in the normal oral mucosa (A-D) and OSCC tumors (E-H) by IHC. (G) Skeletal muscle and (H) neurovascular bundle. N: adjacent non-neoplastic mucosa; T: neoplastic mucosa; SphK2: sphingosine kinase 2; OSCC: oral squamous cell carcinoma; IHC: immunohistochemistry.

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: Expression of SphK2 in the normal oral mucosa (A-D) and OSCC tumors (E-H) by IHC. (G) Skeletal muscle and (H) neurovascular bundle. N: adjacent non-neoplastic mucosa; T: neoplastic mucosa; SphK2: sphingosine kinase 2; OSCC: oral squamous cell carcinoma; IHC: immunohistochemistry.

Techniques: Expressing, Immunohistochemistry

(A) Western blotting for LPP3, SphK2, SGPP1, and actin was performed in tumor tissues (T1, T2, T3, and T4) and adjacent normal tissues (N1, N2, N3, and N4) from the same patients. (B, D, and F) Densitometric analysis was performed with Image J software and the ratio of target protein/beta-actin for each sample is shown with the bar diagram. (C, E, and G) The average of target protein/beta-actin was calculated and values are shown as mean ± standard deviation. A t-test was performed to compare the difference in mean. LPP3: lipid phosphate phosphatase 3; SphK2: sphingosine kinase 2; SGPP1: sphingosine-1-phosphate phosphatase 1; OSCC: oral squamous cell carcinoma.

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: (A) Western blotting for LPP3, SphK2, SGPP1, and actin was performed in tumor tissues (T1, T2, T3, and T4) and adjacent normal tissues (N1, N2, N3, and N4) from the same patients. (B, D, and F) Densitometric analysis was performed with Image J software and the ratio of target protein/beta-actin for each sample is shown with the bar diagram. (C, E, and G) The average of target protein/beta-actin was calculated and values are shown as mean ± standard deviation. A t-test was performed to compare the difference in mean. LPP3: lipid phosphate phosphatase 3; SphK2: sphingosine kinase 2; SGPP1: sphingosine-1-phosphate phosphatase 1; OSCC: oral squamous cell carcinoma.

Techniques: Western Blot, Software, Standard Deviation

Correlation of mRNA expression of S1P-metabolizing enzymes with immune infiltration level in HNSCC (N = 522), HPV-negative (N = 422), and HPV-positive (N = 98) patients was determined using Tumor Immune Estimation Resource 2.0 (TIMER 2.0). Association between mRNA expression of S1P-metabolizing enzymes and tumor-infiltrating immune cells was analyzed by the

Journal: Cureus

Article Title: Downregulation of Lipid Phosphate Phosphatase 3 Correlates With Tumor-Infiltrating Immune Cells in Oral Cancer

doi: 10.7759/cureus.23553

Figure Lengend Snippet: Correlation of mRNA expression of S1P-metabolizing enzymes with immune infiltration level in HNSCC (N = 522), HPV-negative (N = 422), and HPV-positive (N = 98) patients was determined using Tumor Immune Estimation Resource 2.0 (TIMER 2.0). Association between mRNA expression of S1P-metabolizing enzymes and tumor-infiltrating immune cells was analyzed by the "Immune" module of TIMER 2.0 (A). A heat map was generated for showing the positive, negative, or no correlation between gene expression and infiltration level of immune cells (B). Representative dot plots were drafted between PPAP2B expression Log2 TPM (transcript per million) and infiltration level. S1P: sphingosine-1-phosphate; SphK1: sphingosine kinase 1; SphK2: sphingosine kinase 2; SGPP1: sphingosine-1-phosphate phosphatase 1; PLPP3: phospholipid phosphatase 3; HNSCC: head and neck squamous cell carcinoma; HPV: human papillomavirus; PPAP2B: phosphatidic acid phosphatase type 2B.

Techniques: Expressing, Generated, Gene Expression

Review

Structured Review

Images

Similar Products

Image Search Results