s1pr4  (MedChemExpress)

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

Journal: European Journal of Immunology

Article Title: Sphingosine‐1‐phosphate receptor type 4 is critically involved in the regulation of peritoneal B‐1 cell trafficking and distribution in vivo

doi: 10.1002/eji.202350882

Figure Lengend Snippet: Chemotactic response of s1pr 4 –/– peritoneal B cells in vitro (A) Peritoneal B cells were isolated from the PerC of wt and s1pr 4 –/– mice and seeded in the upper chamber of a Transwell migration assay. The lower chambers were filled with different concentrations of S1P alone (solid line) or with 100 ng/mL CXCL13 (dotted line). After incubation, the number of cells that migrated to the lower chamber was quantified using flow cytometry. Migration was computed as the percentage of cells from the upper well. Values represent the mean ± SD of n = 3 per concentration. The results are representative of three independent experiments. (B) Transwell migration of peritoneal B cells to medium, 10 nM S1P, 10 nM S1P + 200 ng/mL CXCL12, and 10 nM S1P + 100 ng/mL CXCL13. Data are shown as individual values obtained from n = 8 donors per group. (C) Expression of CXCR4 and CXCR5 on peritoneal cell populations quantified by flow cytometry ( n = 8). (D) Expression of CXCR4 and CXCR5 on peritoneal B cells that were stimulated with the selective S1PR 4 agonist CYM50308 or selective S1PR 4 antagonist CYM50358 in the presence of S1P. Bars represent the mean values + SD. * p < 0.05; ** p < 0.01; *** p < 0.001 as computed by Student's t ‐test.

Article Snippet: The S1PR 4 ‐specific antagonist CYM50358 (Hycultec) was solubilized according to the recommendations of the manufacturer and used to demonstrate the S1PR 4 ‐specific effects of S1P at the concentrations specified in the results section.

Techniques: In Vitro, Isolation, Transwell Migration Assay, Incubation, Flow Cytometry, Migration, Concentration Assay, Expressing