Journal: Molecular medicine reports

Article Title: Effects of S1PR2 antagonist on blood pressure and angiogenesis imbalance in preeclampsia rats.

doi: 10.3892/mmr.2021.12095

Figure Lengend Snippet: Figure 1. S1PR2 is increased in the serum and placental tissue of PE rats. (A) Measurement of S1P in serum via ELISA. (B) The expression of S1PR1, S1PR2 and S1PR3 in serum of control and PE rats was analyzed by reverse transcription‑quantitative PCR analysis. (C) The effect of S1P on the expression of S1PR2 in the plasma of PE rats was analyzed via ELISA. (D) The effect of S1P on the expression of S1PR2 in the placental tissues of PE rats was analyzed by western blotting. n=3. *P<0.05, **P<0.01 and ***P<0.001 vs. Control group; #P<0.05 and ###P<0.001 vs. Model group; ∆P<0.05 and ∆∆∆P<0.001 vs. S1P group. S1P, sphingosine‑1‑phosphate; S1PR2, sphingosine‑1‑phosphate receptor 2; PE, preeclampsia; ELISA, enzyme‑linked immunosorbent assay.

Article Snippet: The membranes were incubated with primary antibodies against S1PR2 (cat. no. 21180‐1‐AP; 1:1,000; ProteinTech Group, Inc.), VEGF (cat. no. 66828‐1‐AP; 1:1,000; ProteinTech Group, Inc.), Flt‐1 (cat. no. 13687‐1‐AP; 1:1,000; ProteinTech Group, Inc.), endo‐ thelial (e)NOS (cat. no. ab76198; 1:1,000; Abcam) and β‐actin (cat. no. ab8226; 1:1,000; Abcam) overnight at 4 ̊C.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control, Clinical Proteomics, Western Blot

Journal: Molecular medicine reports

Article Title: Effects of S1PR2 antagonist on blood pressure and angiogenesis imbalance in preeclampsia rats.

doi: 10.3892/mmr.2021.12095

Figure Lengend Snippet: Figure 2. Inhibition of S1PR2 with JTE‑013 decreases BP in PE rats. Effect of JTE‑013 on (A) SBP and (B) DBP in PE rats in tail‑cuff measurement. n=3. *P<0.05, **P<0.01 and ***P<0.001 vs. Control group; #P<0.05, ##P<0.01 and ###P<0.001 vs. Model group; ∆P<0.05 vs. Model + S1PR2 antagonist low group. S1PR2, sphingosine‑1‑phosphate receptor 2; PE, preeclampsia; BP, blood pressure; SBP, systolic blood pressure; DBP, diastolic blood pressure.

Article Snippet: The membranes were incubated with primary antibodies against S1PR2 (cat. no. 21180‐1‐AP; 1:1,000; ProteinTech Group, Inc.), VEGF (cat. no. 66828‐1‐AP; 1:1,000; ProteinTech Group, Inc.), Flt‐1 (cat. no. 13687‐1‐AP; 1:1,000; ProteinTech Group, Inc.), endo‐ thelial (e)NOS (cat. no. ab76198; 1:1,000; Abcam) and β‐actin (cat. no. ab8226; 1:1,000; Abcam) overnight at 4 ̊C.

Techniques: Inhibition, Control

Journal: Molecular medicine reports

Article Title: Effects of S1PR2 antagonist on blood pressure and angiogenesis imbalance in preeclampsia rats.

doi: 10.3892/mmr.2021.12095

Figure Lengend Snippet: Figure 3. Effect of JTE‑013 on serum NO and iNOS and the expression of eNOS in placental tissues. (A) Summarized data showing the inhibitory effect of JTE‑013 on serum NO levels in PE rats. (B) Summarized data showing the inhibitory effect of JTE‑013 on serum iNOS levels in PE rats. (C) Summarized data showing that JTE‑013 prevented the decreased expression of eNOS in placental tissues of PE rats. n=3. *P<0.05, **P<0.01 and ***P<0.001 vs. Control group; ##P<0.01 and ###P<0.001 vs. Model group; ∆P<0.05 and ∆∆∆P<0.001 vs. Model + S1PR2 antagonist low group. S1PR2, sphingosine‑1‑phosphate receptor 2; PE, preeclampsia; iNOS, inducible nitric oxide synthase; eNOS, endothelial nitric oxide synthase; NO, nitric oxide.

Article Snippet: The membranes were incubated with primary antibodies against S1PR2 (cat. no. 21180‐1‐AP; 1:1,000; ProteinTech Group, Inc.), VEGF (cat. no. 66828‐1‐AP; 1:1,000; ProteinTech Group, Inc.), Flt‐1 (cat. no. 13687‐1‐AP; 1:1,000; ProteinTech Group, Inc.), endo‐ thelial (e)NOS (cat. no. ab76198; 1:1,000; Abcam) and β‐actin (cat. no. ab8226; 1:1,000; Abcam) overnight at 4 ̊C.

Techniques: Expressing, Control

Journal: Molecular medicine reports

Article Title: Effects of S1PR2 antagonist on blood pressure and angiogenesis imbalance in preeclampsia rats.

doi: 10.3892/mmr.2021.12095

Figure Lengend Snippet: Figure 4. JTE‑013 inhibits the PE model‑induced expression levels of VEGF and Flt‑1 receptor in PE rats. (A) Summarized data showing the inhibitory effect of JTE‑013 on the PE model‑induced mRNA levels of VEGF and Flt‑1 in reverse transcription‑quantitative PCR assay. (B) Representative western blotting images and summarized data showing the inhibitory effect of JTE‑013 on the PE model‑induced protein levels of VEGF and Flt‑1 in the placental tissues of PE rats. n=3. *P<0.05 and ***P<0.001 vs. Control group; #P<0.05, ##P<0.01 and ###P<0.001 vs. Model group; ∆∆P<0.01 and ∆∆∆P<0.001 vs. Model + S1PR2 antagonist low group. S1PR2, sphingosine‑1‑phosphate receptor 2; PE, preeclampsia; VEGF, vascular endothelial growth factor; Flt‑1, fms‑like tyrosine kinase 1.

Article Snippet: The membranes were incubated with primary antibodies against S1PR2 (cat. no. 21180‐1‐AP; 1:1,000; ProteinTech Group, Inc.), VEGF (cat. no. 66828‐1‐AP; 1:1,000; ProteinTech Group, Inc.), Flt‐1 (cat. no. 13687‐1‐AP; 1:1,000; ProteinTech Group, Inc.), endo‐ thelial (e)NOS (cat. no. ab76198; 1:1,000; Abcam) and β‐actin (cat. no. ab8226; 1:1,000; Abcam) overnight at 4 ̊C.

Techniques: Expressing, Western Blot, Control

Journal: Molecular medicine reports

Article Title: Effects of S1PR2 antagonist on blood pressure and angiogenesis imbalance in preeclampsia rats.

doi: 10.3892/mmr.2021.12095

Figure Lengend Snippet: Figure 5. JTE‑013 attenuates pathological changes in placental tissues and decreases inflammation in PE rats. (A) Summarized data showing the inhibitory effect of JTE‑013 on the increased expression of serum TNF‑α, IL‑1β and IL‑6, as determined by an enzyme‑linked immunosorbent assay. (B) Summarized data showing JTE‑013 attenuated the infiltration of inflammatory cells in placental tissues, as detected by hematoxylin and eosin staining (magnification, x100 or 200). n=3. *P<0.05, **P<0.01 and ***P<0.001 vs. Control group; ##P<0.01 and ###P<0.001 vs. Model group; ∆P<0.05 and ∆∆P<0.01 vs. Model + S1PR2 antagonist low group. PE, preeclampsia; TNF‑α, tumor necrosis factor‑α; IL‑, interleukin; S1PR2, sphingosine‑1‑phosphate receptor 2; lab, labyrinth; JZ, junctional zone; de, decidua.

Article Snippet: The membranes were incubated with primary antibodies against S1PR2 (cat. no. 21180‐1‐AP; 1:1,000; ProteinTech Group, Inc.), VEGF (cat. no. 66828‐1‐AP; 1:1,000; ProteinTech Group, Inc.), Flt‐1 (cat. no. 13687‐1‐AP; 1:1,000; ProteinTech Group, Inc.), endo‐ thelial (e)NOS (cat. no. ab76198; 1:1,000; Abcam) and β‐actin (cat. no. ab8226; 1:1,000; Abcam) overnight at 4 ̊C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Control

Journal: Mediators of Inflammation

Article Title: Sphingosine-1-Phosphate/Sphingosine-1-Phosphate Receptor 2 Axis Can Promote Mouse and Human Primary Mast Cell Angiogenic Potential through Upregulation of Vascular Endothelial Growth Factor-A and Matrix Metalloproteinase-2

doi: 10.1155/2016/1503206

Figure Lengend Snippet: Role of S1P/S1PR2 signaling in VEGF-A secretion. (a) Blood was collected from allergenically challenged WT (open bar, n = 5-6 mice) and S1PR2-null (black bar, n = 5-6 mice) mice, euthanized 2 hours after Ag challenge, and serum VEGF-A levels were measured in duplicate determination for each animal. (b) Bone marrow-derived mast cells (BMMC, three independent populations) from both genotypes were stimulated for 24 hours with vehicle (DMSO/PBS/4 mg/mL fatty acid-free BSA), S1P (100 nM), IgE/Ag, or ionomycin (Iono) and VEGF-A was measured in MC supernatants, in duplicate determinations. Error bars show standard error of means. ∗ p < 0.05, ∗∗ p < 0.005.

Article Snippet: S1PR2 knockout mice (Taconic Biosciences, Inc., Hudson, NY) and corresponding wild-type (WT) mice were on a mixed 129/SvEV-C57BL/6 background.

Techniques: Derivative Assay

Journal: Mediators of Inflammation

Article Title: Sphingosine-1-Phosphate/Sphingosine-1-Phosphate Receptor 2 Axis Can Promote Mouse and Human Primary Mast Cell Angiogenic Potential through Upregulation of Vascular Endothelial Growth Factor-A and Matrix Metalloproteinase-2

doi: 10.1155/2016/1503206

Figure Lengend Snippet: Human MC secrete proangiogenic factors upon exposure to S1P in a S1PR2-dependent manner. (a) VEGF-A levels were measured in the supernatants of activated human MC in the absence (open bars, vehicle (DMSO/PBS/4 mg/mL fatty acid-free BSA)) or presence of a 30-minute pretreatment with CYM-5442 (filled bars, S1PR1 agonist, 1 μ M) or with JTE-013 (gray bars, S1PR2 antagonist, 1 μ M), followed by the indicated stimuli similar to . (b) Matrix metalloproteinase- (MMP-) 2 levels were measured in supernatants of MC activated exactly as described in . All supernatants were collected 24 hours after stimulation. Activation experiments were conducted using five independent human skin MC populations generated from five donors. Activation was conducted in triplicate determinations and measurements were conducted in duplicate determinations for each individual determination. When reaching significance, statistics are indicated in each figure. Error bars show standard error of means. ∗∗∗ p < 0.0001.

Article Snippet: S1PR2 knockout mice (Taconic Biosciences, Inc., Hudson, NY) and corresponding wild-type (WT) mice were on a mixed 129/SvEV-C57BL/6 background.

Techniques: Activation Assay, Generated

Journal: Acta Histochemica et Cytochemica

Article Title: Effect of Hepatic Lipid Overload on Accelerated Hepatocyte Proliferation Promoted by HGF Expression via the SphK1/S1PR2 Pathway in MCD-diet Mouse Partial Hepatectomy

doi: 10.1267/ahc.24-00046

Figure Lengend Snippet: SphK1/S1P signaling in mouse liver during regeneration. qPCR analysis of ( A ) SphK1 , ( B ) SphK2 , ( C ) S1PR3 , and ( D ) ABCC1 mRNA expression in the liver of chow- and MCD-diet fed mice after PHx. ( E ) Immunofluorescence analysis of S1PR2 (green) and DAPI (blue) in mouse liver after PHx. Bar = 20 μm; 40× objective lenses were used. ( F ) Quantitative analysis of S1PR2-positive cells in the liver of chow- and MCD-diet fed mice after PHx. Data represent the mean ± SEM for 6–8 mice per group. Asterisks indicate statistically significant differences (*p < 0.05, **p < 0.01 and ***p < 0.001).

Article Snippet: S1PR2-expressing cells were stained with rabbit polyclonal antibody against S1PR2 (Novus Biologicals) followed by treatment with Alexa-633 conjugated goat anti-Rabbit IgG.

Techniques: Expressing, Immunofluorescence

Journal: Acta Histochemica et Cytochemica

Article Title: Effect of Hepatic Lipid Overload on Accelerated Hepatocyte Proliferation Promoted by HGF Expression via the SphK1/S1PR2 Pathway in MCD-diet Mouse Partial Hepatectomy

doi: 10.1267/ahc.24-00046

Figure Lengend Snippet: Expression of S1PR2 in mouse liver. Double immunofluorescence analysis of S1PR2 (green) and HNF-4α (red staining in upper panel), F4/80 (red staining in middle panel), Ly6G (red staining in lower panel), and DAPI (blue) in the liver of MCD-diet fed mice at 12hr after PHx. Arrows indicate double-positive cells. Bar = 20 μm.

Article Snippet: S1PR2-expressing cells were stained with rabbit polyclonal antibody against S1PR2 (Novus Biologicals) followed by treatment with Alexa-633 conjugated goat anti-Rabbit IgG.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Acta Histochemica et Cytochemica

Article Title: Effect of Hepatic Lipid Overload on Accelerated Hepatocyte Proliferation Promoted by HGF Expression via the SphK1/S1PR2 Pathway in MCD-diet Mouse Partial Hepatectomy

doi: 10.1267/ahc.24-00046

Figure Lengend Snippet: HGF and FGF2 mRNA expression in S1PR2-positive cells after PHx. ( A ) Macrophages (F4/80) and neutrophils (Ly6G) were isolated from the liver of chow- and MCD-diet fed mice at 0, 6, and 12 hr after PHx. The percentage of macrophages ( B ) and neutrophils ( C ) in the liver of chow- and MCD-diet fed mice. HGF and FGF2 mRNA expression levels in ( D , F ) macrophages and ( E , G ) neutrophils isolated from the liver of chow- and MCD-diet fed mice at 0, 6 and 12 hr after PHx. Data represent the mean ± SEM for 3 mice per group. Asterisks indicate statistically significant differences (*p < 0.05, **p < 0.01 and ***p < 0.001).

Article Snippet: S1PR2-expressing cells were stained with rabbit polyclonal antibody against S1PR2 (Novus Biologicals) followed by treatment with Alexa-633 conjugated goat anti-Rabbit IgG.

Techniques: Expressing, Isolation