Journal: Journal of Inflammation (London, England)

Article Title: Enhanced inflammasome activation and reduced sphingosine-1 phosphate S1P signalling in a respiratory mucoobstructive disease model

doi: 10.1186/s12950-020-00248-2

Figure Lengend Snippet: Colocalization and correlation of SPHK1 expression with SPNS2. a Representative confocal images of a control animal bronchiole showing colocalization of SPHK1 (red) and SPNS2 (green). Yellow is merged color of red and green. Blue is DAPI. Scale bars are in micrometers. b Positive Spearman’s correlation between MFI of SPHK1 and SPNS2 in both unobstructed (red; r = 0.640; p < 0.001; n = 62 bronchioles from 7 mice) and mucus obstructed (green; r = 0.745; p < 0.001; n = 42 bronchioles from 4 mice)

Article Snippet: Primary antibodies included rabbit polyclonal to SPHK1 and SPHK2 (Bs-2652R and Bs-2653R, Bioss, Woburn, MA, USA, both 1:50), NLRP3 and IL-1β (sc-66,846 and sc-7884, Santa Cruz, Dallas, TX, USA, both 1:40), GLUT1 (Glucose Transporter 1, ab652, Abcam, Cambridge, UK, 1:100), goat polyclonal antibody to SPNS2 (sc-165,572, Santa Cruz, 1:40), TNFα (sc-1348, Santa Cruz, 1:50), myeloperoxidase (#2141, Osenses, Adelaide, SA, Australia, 1:100), and a mouse monoclonal antibody to neutrophil elastase (M0752, Dako GmbH, Jena, Germany, 1:100).

Techniques: Expressing

Journal: Journal of Inflammation (London, England)

Article Title: Enhanced inflammasome activation and reduced sphingosine-1 phosphate S1P signalling in a respiratory mucoobstructive disease model

doi: 10.1186/s12950-020-00248-2

Figure Lengend Snippet: Hypothetical model of NLRP3 inflammasome activation and S1P signalling dysregulation in mucus-obstructed bronchioles. Unobstructed bronchioles (AIR): NLRP3 is mostly in monomeric state, only background levels of cleaved IL-1β are detected; SPHK1 generates cytosolic S1P, part of which is exported via SPNS2 for extracellular (inside-out) signalling via S1PRs; SPHK2 generates S1P mostly for intracellular signalling. Obstructed bronchioles (MUCUS): Increased oligomerization of NLRP3 (inflammasome activation) in either epithelium or infiltrating leucocytes results in IL-1β cleavage and increased luminal specks of NLRP3/IL-1β; decrease of SPNS2 and SPHK2 results in reduction of extra- and intracellular S1P signalling

Article Snippet: Primary antibodies included rabbit polyclonal to SPHK1 and SPHK2 (Bs-2652R and Bs-2653R, Bioss, Woburn, MA, USA, both 1:50), NLRP3 and IL-1β (sc-66,846 and sc-7884, Santa Cruz, Dallas, TX, USA, both 1:40), GLUT1 (Glucose Transporter 1, ab652, Abcam, Cambridge, UK, 1:100), goat polyclonal antibody to SPNS2 (sc-165,572, Santa Cruz, 1:40), TNFα (sc-1348, Santa Cruz, 1:50), myeloperoxidase (#2141, Osenses, Adelaide, SA, Australia, 1:100), and a mouse monoclonal antibody to neutrophil elastase (M0752, Dako GmbH, Jena, Germany, 1:100).

Techniques: Activation Assay

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 3. Correlation of SPHK1 gene expression with various clinicopathological characteristics in breast tumor tissues. (A) pN Stage (pN0 + pNx- [Nodal Negative], pN1 + pN2-[Nodal Positive]) (B) pTNM Stage (pTNM I + II- [early stage], pTNM III + IV- [late stage]) (C) Correlation with Ki67.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Gene Expression

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 4. Protein expression of SPHK1 and CERK in breast cancer patients. (A) Representative blots in adjacent normal (N) and tumor (T) tissues, (B) Densitometric analysis of SPHK1 and (C) CERK levels in adjacent normal and tumor tissues.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Expressing

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 6. Network-based analysis of interaction network corresponding to candidate genes. (A) The node degree distribution of PPI network. The number of genes is plotted as a function of their degree reflecting a power-law like distribution. The red line corresponds to a power-law distribution. (B) Sub-network of candidate genes. The large-sized black-colored nodes represent the candidate genes (ABCC1, ABCG2, CERK, MMP-2, MMP-9, and SPHK1), while the small gray color nodes represent the corresponding interactor genes. Interactions are represented in gray color and the depth of color represents the strength of correlations. (C) Shortest path lengths among candidate genes. Heatmap of shortest path length among the candidate genes, where the values represent the number of shortest paths between any pair.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 8. Correlation of CERK with (A) MMP-2, (B) MMP-9 and SPHK1 with (C) MMP-2, (D) MMP-9 in local cohort and CERK with (E) MMP-2, (F) MMP-9 and SPHK1 with (G) MMP-2, (H) MMP-9 in TCGA cohort.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 10. Correlation of CERK with (A) ABCC1, (B) ABCG2 and SPHK1 with (C) ABCC1, (D) ABCG2 in local cohort and CERK with (E) ABCC1, (F) ABCG2 and SPHK1 with (G) ABCC1, (H) ABCG2 in TCGA cohort.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 11. Expression of SPNS2 in TCGA cohort. (A) Relative gene expression between adjacent normal and tumor tissue, (B) Correlation with SPHK1.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Expressing, Gene Expression

Journal: Molecular Medicine Reports

Article Title: Ginsenoside Rg1 exerts anti-apoptotic effects on non-alcoholic fatty liver cells by downregulating the expression of SGPL1

doi: 10.3892/mmr.2022.12694

Figure Lengend Snippet: Effect of Rg1 on sphingolipid pathway proteins. (A) SPHK1, SGPL1, p-Akt, p-Erk1/2, Akt and Erk1/2 expression levels were detected in HHL-5 cells treated with Rg1 (0.2, 0.4 and 0.6 mM) after exposure to MCE (1%) by western blotting. (B) SPHK1 and SGPL1 mRNA expression levels were detected in HHL-5 cells using reverse transcription-quantitative PCR. (C) SPHK1 and SGPL1 protein expression levels were normalized to actin expression, and p-Akt and p-Erk1/2 protein expression levels were normalized to Akt or Erk1/2 using ImageJ software. *P<0.05, **P<0.01 and ***P<0.001; MCE, medium- and long-chain fat emulsion; Mod, model; p-, phosphorylated; Rec, recovery; SGPL1, sphingosine-1-phosphate lyase 1; SPHK1, sphingosine kinase 1; Rg1, ginsenoside Rg1.

Article Snippet: The antibody against SPHK1 (AF5536) was obtained from R&D Systems. β-actin antibody (1:8,000; cat. no. A1978; MilliporeSigma) was used as a loading control.

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Software, Emulsion

Journal: Molecular Medicine Reports

Article Title: Ginsenoside Rg1 exerts anti-apoptotic effects on non-alcoholic fatty liver cells by downregulating the expression of SGPL1

doi: 10.3892/mmr.2022.12694

Figure Lengend Snippet: Molecular mechanism underlying the inhibitory effects of Rg1 on apoptosis in steatotic HHL-5 hepatocytes. MCE, medium- and long-chain fat emulsion; SPHK1, sphingosine kinase 1; SGPL1, sphingosine-1-phosphate lyase 1; S1P, sphingosine-1-phosphate; Mt, mitochondria; Rg1, ginsenoside Rg1.

Article Snippet: The antibody against SPHK1 (AF5536) was obtained from R&D Systems. β-actin antibody (1:8,000; cat. no. A1978; MilliporeSigma) was used as a loading control.

Techniques: Emulsion

Journal: European Journal of Histochemistry : EJH

Article Title: Interaction between sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in experimental intestinal fibrosis. An in vivo immunohistochemical study

doi: 10.4081/ejh.2018.2956

Figure Lengend Snippet: Antibodies used with their sources and dilutions together with the positive controls.

Article Snippet: SPHK1 , (PA1996) BosterBio, Pleasanton, CA, USA , 1:100 , Human kidney cancer.

Techniques:

Journal: European Journal of Histochemistry : EJH

Article Title: Interaction between sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in experimental intestinal fibrosis. An in vivo immunohistochemical study

doi: 10.4081/ejh.2018.2956

Figure Lengend Snippet: Immunohistochemistry (original magnification 10x). The immunopositivity of SPHK1, RhoA and PI3K is prevalent in DSS mice (B,E,H) respect to control (A,D,G). In particular, in DSS mice, SPHK1 expression is predominantly localized in mucosa layer and in submucosa of colon (B). In the same group RhoA and PI3K staining results highly expressed (E,H) in mucosa, especially in the ephitelium (supplementary Figure 1), but also in submucosa and in serosa.

Article Snippet: SPHK1 , (PA1996) BosterBio, Pleasanton, CA, USA , 1:100 , Human kidney cancer.

Techniques: Immunohistochemistry, Control, Expressing, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: Interaction between sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in experimental intestinal fibrosis. An in vivo immunohistochemical study

doi: 10.4081/ejh.2018.2956

Figure Lengend Snippet: Schematic representation of TGF-β dependent signaling implicated in fibrosis process. TGF-β upon biding its receptors, activates both canonical Smads pathway and noncanonical pathway (SPHK1/S1P/mTOR), inducing transcription of pro-fibrotic target genes.

Article Snippet: SPHK1 , (PA1996) BosterBio, Pleasanton, CA, USA , 1:100 , Human kidney cancer.

Techniques: