t22703  (Topscience Co Ltd)

Journal: iScience

Article Title: Broad and diverse roles of sphingosine-1-phosphate/sphingosine-1-phosphate receptors in the prostate.

doi: 10.1016/j.isci.2024.111290

Figure Lengend Snippet: Figure 8. S1P/S1PR2/S1PR3 induced contraction of WPMY-1 cells and potentiated PE-mediated contraction of human prostate strips via RhoA/ROCK pathway (A) The effect of S1PRs knockdown and overexpression on contraction of WPMY-1 cells. (B) Statistical analysis of the percentages of the area of collagen gels. (C) Western blot assay of pathway-related proteins. (D) The effect of SEW2871, CYM5520 and CYM5541 on contraction of human prostate tissue strips. (E) Statistical analysis of tension (KCl = 100%). (F) Statistical analysis of time to 50% maximal 105 M PE induced contraction. Data are represented as M ± SD. *p < 0.05, **p < 0.01 and ns means no significant difference.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies See Tables 3 and 4 This paper N/A Biological samples Human bladder, kidney, liver, lung, lymph node, prostate, sperm duct, spleen, testis, ureter, vessel tissues and serum Zhongnan Hospital, Wuhan University N/A Rat bladder, brain, heart, kidney, liver, lung, penis, prostate, spleen, testis and vessel tissues Normal male Sprague-Dawley rats N/A Chemicals, peptides, and recombinant proteins S1P Sigma Cat. 73914 U0126 MCE Cat. HY-12031A MK2206 MCE Cat. HY-108232 SEW2871 MCE Cat. HY-W008947 CYM5520 Topscience Cat. T22703 CYM5541 MCE Cat. HY-101419 W-146 MCE Cat. HY-101395 TY-52156 MCE Cat. HY-19736 Critical commercial assays Hipure Total RNA Mini Kit Magen Cat. R4111-03 ABScript II RT Master Mix for qPCR Abclonal Cat. RK20402 23 Universal SYBR Green Fast qPCR Mix Abclonal Cat. RK21203 Cell cycle staining kit Multi Sciences Cat. CCS012 Annexin V-FITC/PI apoptosis kit Multi Sciences Cat. AP101-100-kit Cell counting kit-8 Meilun Biotechnology Cat. MA0218-L Cell Contraction Assay Kit Cell Biolabs Cat. CBA-201 Human S1P ELISA kit Meimian Cat. MM-91866O1 rat IL-6 ELISA kit Meimian Cat. MM-0190R2 rat IL-8 ELISA kit Meimian Cat. 0175R2 rat TNF-aELISA kit Meimian Cat. MM-0180R2 Experimental models: Cell lines RWPE-1 ATCC Cat. CRL11609 WPMY-1 Stem Cell Bank Cat. GNHu36 Experimental models: Organisms/strains Sprague-Dawley rat This paper N/A Oligonucleotides si-RNA, see Table 5 Genepharma N/A Primer for RT-qPCR, see Tables 1 and 2 This paper N/A Recombinant DNA S1PR1, S1PR2, S1PR3 Fenghui Cat. JY3307, Cat. JY4554, Cat. JY4555 Software and algorithms Adobe Illustrator Adobe https://www.adobe.com/ ImageJ NIH https://imagej.nih.gov/ij/ Prism Graphpad https://www.graphpad.com e1 iScience 27, 111290, December 20, 2024

Techniques: Knockdown, Over Expression, Western Blot

Journal: iScience

Article Title: Broad and diverse roles of sphingosine-1-phosphate/sphingosine-1-phosphate receptors in the prostate.

doi: 10.1016/j.isci.2024.111290

Figure Lengend Snippet: Figure 8. S1P/S1PR2/S1PR3 induced contraction of WPMY-1 cells and potentiated PE-mediated contraction of human prostate strips via RhoA/ROCK pathway (A) The effect of S1PRs knockdown and overexpression on contraction of WPMY-1 cells. (B) Statistical analysis of the percentages of the area of collagen gels. (C) Western blot assay of pathway-related proteins. (D) The effect of SEW2871, CYM5520 and CYM5541 on contraction of human prostate tissue strips. (E) Statistical analysis of tension (KCl = 100%). (F) Statistical analysis of time to 50% maximal 105 M PE induced contraction. Data are represented as M ± SD. *p < 0.05, **p < 0.01 and ns means no significant difference.

Article Snippet: For human prostate, strips were washed and incubated with S1P, SEW2871, CYM5520 (S1PR2 agonist, Topscience, Cat. T22703) and CYM5541 for 20 min, respectively, after stimulating by 10 4 M PE.

Techniques: Knockdown, Over Expression, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Spinster homolog 2/S1P signaling ameliorates macrophage inflammatory response to bacterial infections by balancing PGE 2 production

doi: 10.1186/s12964-024-01851-z

Figure Lengend Snippet: Spns2/S1P signaling impacts mitochondrial functions through coordinated activation of multiple S1P receptors (A) All five S1PRs are transcriptionally detectable in PMs. N = 3 biological replicates. (B) Inhibition of individual S1PRs in WT PMs increases the gene expression of Ptges , while activation of S1PR2 and S1PR4 in Spns2 −/− PMs may slightly reduce Ptges expression. N = 4 biological replicates. (C) In WT PMs, blocking individual S1PRs reduces the fluorescence intensity of MitoTracker™ Red, indicating diminished ΔΨm. (D) S1PR3 blockade significantly increases the fluorescence intensity of CytoFix™ MitoRed, indicating increased mitochondrial mass, while blockade of other receptors may cause a slight increase in mitochondrial mass. (E) All treatments result in reduced average Δψm (calculated by the ratio of MT/CF) in WT PMs. N = 3 biological replicates ( C to E ). ( F , G ) In Spns2 −/− PMs, activation of S1PR3 and S1PR5 increases MitoTracker™ Red fluorescence intensity (F) , but none of the treatments affect mitochondrial mass (G) . (H) Only S1PR3 activation partially restores the average Δψm in Spns2 −/− PMs. N = 3 biological replicates ( F to H ). (I) All the antagonists increase mtROS generation in WT PMs, especially with S1PR2 and S1PR4 blockade. N = 3 biological replicates. (J) In Spns2 −/− PMs, activation of S1PR2 and S1PR4 attenuates oxidative stress. N = 3 biological replicates. Data are presented as mean ± s.e.m. P values were determined by unpaired t -test (A) and one-way ANOVA with Sidak’s correction for multiple comparisons ( B to J ). * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant

Article Snippet: PMs were pre-treated with the following compounds for 1-h before analysis, unless otherwise stated: 5 µM Spns2 inhibitor SLF1081851 (MedChemExpress, HY-149004), 1 µM S1P, 100 nM PF-04418948, 100 nM ONO-AE3-208, 200 nM PGE 2 (TargetMOI, T5014), 2 µM S1PR1 antagonist W146 (Cayman Chemical, 10009109), 2 µM S1PR2 antagonist JTE013 (Cayman Chemical, 10009458), 2 µM S1PR3 antagonist TY-52,156 (TargetMOI, T17183), 2 µM S1PR4 antagonist CYM50358 (Tocris Bioscience, 4679), 5 µM S1PR1 agonist CYM5442 (TargetMOI, T2026), 5 µM S1PR2 agonist CYM5520 (TargetMOI, T22703), 5 µM S1PR3 agonist CYM5541 (TargetMOI, T3961), 5 µM S1PR4 agonist CYM50260 (TargetMOI, T15031), and 50 nM S1PR5 agonist A971432 (Cayman Chemical, 25326).

Techniques: Activation Assay, Inhibition, Expressing, Blocking Assay, Fluorescence