Journal: PLoS ONE

Article Title: Knock Out of S1P3 Receptor Signaling Attenuates Inflammation and Fibrosis in Bleomycin-Induced Lung Injury Mice Model

doi: 10.1371/journal.pone.0106792

Figure Lengend Snippet: S1P concentration in BALF collected on the seventh day after intratracheal administration of bleomycin or non-treated control. There were no significant differences in S1P concentration in BALF between wild-type (WT) and knockout (KO) mice at baseline or on the seventh day after bleomycin administration, nor were there significant differences in the S1P concentrations in BALF collected before and after bleomycin administration in WT (baseline vs. seventh day; p = 0.14) or KO (baseline vs. seventh day; p = 0.50) mice.

Article Snippet: ELISA kits for TGF-β1 and MCP-1 were provided by eBioscience (San Diego, CA, USA), for CTGF by Uscn Life Science (Houston, TX, USA) and for S1P by Echelon Biosciences Inc. (Salt Lake City, UT, USA).

Techniques: Concentration Assay, Knock-Out

Journal: iScience

Article Title: Mouse phospholipid phosphatase 6 regulates dendritic cell cholesterol, macropinocytosis, and allergen sensitization

doi: 10.1016/j.isci.2022.105185

Figure Lengend Snippet: Murine Plpp6 gene expression and activity is regulated during inflammation (A) PLPP6 converts presqualene diphosphate (PSDP) into presqualene monophosphate (PSMP) upon cell activation. (B) Phosphate release from PSDP, sphingosine-1-phosphate (S1P), farnesyl diphosphate (FDP), phosphatidic acid (PA), and lysophosphatidic acid (LPA) exposed to recombinant murine PLPP6 (see ); experiments were performed 4 times for a total of n = 3 to 4. (C) Allergic lung inflammation model with house dust mite (HDM) exposure (see ). (D) Lung Plpp6 expression in WT mice on day 16 of HDM protocol; experiments were performed 2 times for a total of n = 4 to 7; data represent fold change relative to baseline expression. (E) Extracted ion chromatogram (MS-XIC) and MS/MS assignment of isolated PSMP ([M−H] = 505.35). (F) PSMP content in lungs from WT and Plpp6 −/− mice on day 16 of HDM protocol; results are expressed as fold change to the mean value of PSMP in the naive group of the same genotype; experiments were performed 3 times for a total of n = 3. ∗ p < 0.05 and ∗∗ p < 0.01 comparing S1P, FDP, PA, LPA to PSDP by one-way ANOVA and Tukey test for multiple comparisons. ## p < 0.01 comparing inflammation to baseline by unpaired nonparametric Mann-Whitney test. Bars represent median with interquartile range. See also Figure S1 .

Article Snippet: Recombinant mPlpp6 (2μg) was added to mixed micelles of select phosphorylated lipids (presqualene diphosphate (PSDP) (isolated as in ( )), sphingosine-1-phosphate (S1P) (Cayman), farnesyl diphosphate (FDP) (Echelon), phosphatidic acid (PA) (Cayman) and lysophosphatidic acid (LPA) (Cayman)) (20μM) for 30 min at 37°C with gentle mixing.

Techniques: Expressing, Activity Assay, Activation Assay, Recombinant, Tandem Mass Spectroscopy, Isolation, MANN-WHITNEY

Journal: iScience

Article Title: Mouse phospholipid phosphatase 6 regulates dendritic cell cholesterol, macropinocytosis, and allergen sensitization

doi: 10.1016/j.isci.2022.105185

Figure Lengend Snippet:

Article Snippet: Recombinant mPlpp6 (2μg) was added to mixed micelles of select phosphorylated lipids (presqualene diphosphate (PSDP) (isolated as in ( )), sphingosine-1-phosphate (S1P) (Cayman), farnesyl diphosphate (FDP) (Echelon), phosphatidic acid (PA) (Cayman) and lysophosphatidic acid (LPA) (Cayman)) (20μM) for 30 min at 37°C with gentle mixing.

Techniques: Clone Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Software, Staining, Cell Stimulation, Antibody Labeling

Journal: International Journal of Molecular Sciences

Article Title: Effect of Empagliflozin on Sphingolipid Catabolism in Diabetic and Hypertensive Rats

doi: 10.3390/ijms23052883

Figure Lengend Snippet: The effect of empagliflozin on sphingosine-1-phosphate content in plasma, kidney, heart, and liver of diabetic rats. Control (C), diabetic (D), and diabetic rats treated with empagliflozin (D+E). Each bar represents the mean ± SE of n = 10. * p < 0.05 when compared with control; δ p < 0.05 when compared with diabetic rats.

Article Snippet: Sphingosine and sphingosine-1 phosphate contents were determined using a commercial ELISA Kit, according to the manufacturer’s instruction (OKEH02615, Aviva Systems Biology, San Diego, CA, and TDS K1900, Echelon Biosciences, Salt Lake City, UT, USA, respectively) [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Effect of Empagliflozin on Sphingolipid Catabolism in Diabetic and Hypertensive Rats

doi: 10.3390/ijms23052883

Figure Lengend Snippet: The effect of empagliflozin on sphingosine-1-phosphate content in plasma, kidney, heart, and liver of hypertensive rats. Normotensive (Sham), hypertensive (Ang II-induced hypertensive rats), and empagliflozin-treated hypertensive rats (Ang II + E). Each bar represents the mean ± SE of n = 10. * p < 0.05 when compared with sham; δ p < 0.05 when compared with hypertensive rats.

Article Snippet: Sphingosine and sphingosine-1 phosphate contents were determined using a commercial ELISA Kit, according to the manufacturer’s instruction (OKEH02615, Aviva Systems Biology, San Diego, CA, and TDS K1900, Echelon Biosciences, Salt Lake City, UT, USA, respectively) [ ].

Techniques:

Journal: Development (Cambridge, England)

Article Title: Evidence that ABC transporter-mediated autocrine export of an eicosanoid signaling molecule enhances germ cell chemotaxis in the colonial tunicate Botryllus schlosseri

doi: 10.1242/dev.184663

Figure Lengend Snippet: Migration of integrin-alpha-6-positive germ cell precursors towards low concentrations of Sphingosine-1-Phosphate (S1P) depends on ABC transporter activity. (A) Representative example of FISH showing expression of abcb1 in vasa+ cells. Dashed line outlines the germ cell niche on the primary bud. All vasa+ cells (red) co-express abcb1 (green). The white arrow indicates a cluster of small germline stem cells, the blue arrow indicates a maturing oocyte. Red and green channels are shown individually with nuclear counterstaining (Hoechst 3342, blue), and merged images on the right show co-expression of both genes (yellow). Gray box indicates magnified region of the merged image. Scale bars: 20 μm. (B) Transwell migration assay of IA6+ cells in response to different concentrations of S1P, with or without inhibitors of ABC transporters. Sphingosine-1-Phosphate (0.2-2 μM), ABCB1-inhibitor (10 μM CP 1000356 hydrochloride), ABCC1 inhibitor (10 μM probenecid), or ABCB1 and ABCC1 inhibitor (10 μM reversan) were added to the bottom wells where indicated. Control wells contain only migration medium. IA6+ cells were added to the upper chamber of an 8 μm transwell filter coated with laminin; after 2 h, migrated cells in the lower chamber were counted. Data are expressed as fold changes of the numbers of migrated cells, normalized to controls (n=4). Statistical analysis was performed using a paired two-tailed Student's t-test.

Article Snippet: Sphingosine-1-Phosphate (0.2-2 μM, Echelon), 12(S)-Hydroxy-(5Z,8Z,10E,14Z)-eicosatetraenoic acid (80 nM, Sigma-Aldrich), 10 μM CP 1000356 hydrochloride (ABCB1 inhibitor), 10 μM probenecid (ABCC inhibitor), 10 μM reversan (ABCB1 and ABCC1 inhibitor), 10 μM AACOCF3 (inhibitor of phospholipase A2), 0.5 μM 2-TEDC (inhibitor of 5-, 12- and 15-lipoxygenase), 10 μM zileuton (inhibitor of 5-lipoxygenase), 1 μM BAY-u 9773 (Cysteinyl leukotriene receptor antagonist), 10 μM (S)-(+)-ibuprofen (non-selective but stronger inhibition of Cox-1), 1 mM naproxen (non-selective Cox inhibitor), 1 mM indomethacin (non-selective but stronger inhibition of Cox-1) and 1 mM SC 236 (Cox-2 inhibitor) (all from Tocris) were added to the migration medium in the bottom chamber as indicated.

Techniques: Migration, Activity Assay, Expressing, Transwell Migration Assay, Two Tailed Test