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Cay10444

Manufactured by Cayman Chemical
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Sourced in United States

CAY10444 is a chemical compound produced by Cayman Chemical. It is commonly used in scientific research and laboratory settings. The core function of CAY10444 is to serve as a research tool, but a detailed description of its intended use cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Jte 013, by Cayman Chemical (12 mentions) Sphingosine 1 phosphate (s1p), by Cayman Chemical (4 mentions) Sew2871, by Bio-Techne (3 mentions) Fetal bovine serum (fbs), by Harvard Bioscience (3 mentions) Sew2871, by Cayman Chemical (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Sphingosine 1 phosphate (s1p), by Avanti Polar Lipids (2 mentions) Cym5541, by Bio-Techne (2 mentions) Histopaque 1077, by Merck Group (2 mentions) Anti gapdh, by Santa Cruz Biotechnology (2 mentions) Sphingosine 1 phosphate (s1p), by Merck Group (2 mentions) Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions) Nsc23766, by Cayman Chemical (1 mentions) Bovine fatty acid free albumin, by Merck Group (1 mentions) U0126, by R&D Systems (1 mentions) Ly294002, by R&D Systems (1 mentions) Pd98059, by R&D Systems (1 mentions) Anti f4 80 antibody, by BD (1 mentions) Anti phospho akt, by Cell Signaling Technology (1 mentions) Anti phospho p42 p44 mapk, by Cell Signaling Technology (1 mentions)

26 protocols using cay10444

1

Effects of S1P and S1PR3 Antagonist on SPK1

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The effects were compared of exogenous S1P and/or a S1PR3 antagonist (CAY10444) on SPK1 expression in TKE2 corneal epithelial cells or ocular fibroblasts. Both of these cell types express S1PR3 as detected by immunohistochemistry. They were grown to confluence in a 60 mm culture dish. Then the cells were incubated in a serum-free medium for another 16 h. Cells were cultured under the following conditions for another 24 h under one of the following four different conditions: (a) S1P (200 nM) (Cayman Chemical, Ann Arbor, MI, USA); (b) CAY10444 (a selective S1PR3 antagonist, 100 μM, (Cayman Chemical); (c) or both S1P and CAY10444 at the aforementioned concentrations. (d) serum- free culture served as the control. Ten dishes were prepared for each culture condition. RNA was extracted and processed for real-time qRT-PCR for VEGF-A. Mann–Whitney U test was used for data analysis.
Yasuda S., Sumioka T., Iwanishi H., Okada Y., Miyajima M., Ichikawa K., Reinach P.S, & Saika S. (2020). Loss of sphingosine 1-phosphate receptor 3 gene function impairs injury-induced stromal angiogenesis in mouse cornea. Laboratory Investigation; a Journal of Technical Methods and Pathology, 101(2), 245-257.
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2

Murine Bone Marrow Stromal Cell Isolation

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The BM cells were flushed from the femur and tibia of 20 g ICR mice and were filtered through a 70-μm nylon mesh. Then, cells were washed by phosphate-buffered saline (PBS) containing 2% fetal bovine serum (FBS) and cultured with α-minimum essential medium (MEM) (Invitrogen) containing 20% FBS. The medium was all changed on the 2nd day and half changed on the 3rd day. After the first cell passage, the cells were cultured in α-MEM containing 15% FBS. Passages of 3–6 BMSCs were used for experiments. If not specifically stated, cells were collected after treatment with S1P (Cay62570-1, Biomol, Tebu, France) for 24 h, and CAY10444 (S1PR3 inhibitor; 10,005,033, Cayman Chemical, Ann Arbor, MI), MLS-573151 (Cdc42 inhibitor; 0,453,647-3, Cayman Chemical), and NSC-23766 (Rac1 inhibitor; 0,441,977-5, Cayman Chemical) were added 1 h before the addition of S1P.
Ma Y., Chang N., Liu Y., Liu F., Dong C., Hou L., Qi C., Yang L, & Li L. (2021). Silencing IQGAP1 alleviates hepatic fibrogenesis via blocking bone marrow mesenchymal stromal cell recruitment to fibrotic liver. Molecular Therapy. Nucleic Acids, 27, 471-483.
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3

Sphingosine-1-Phosphate Signaling Assays

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Sphingosine-1-Phosphate (d18:1; Lipid Maps LMSP01050001) was purchased from Avanti Polar Lipids and Sigma; bovine fatty acid free albumin was from Sigma; W146, CAY10444 and ML-031 were from Cayman Chemical; SEW2871 and CYM5541 were from Tocris Bioscience; LY294002, U0126 and PD98059 were from R&D systems.
Ruiz M., Okada H, & Dahlbäck B. (2017). HDL-associated ApoM is anti-apoptotic by delivering sphingosine 1-phosphate to S1P1 & S1P3 receptors on vascular endothelium. Lipids in Health and Disease, 16, 36.
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4

STZ-Induced Liver Ischemia-Reperfusion Injury

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Streptozotocin (STZ; 40 mg/kg) or vehicle control (sodium citrate buffer) was injected intraperitoneally (IP) into separate groups of 6‐week‐old mice for 5 consecutive days. Mice were anesthetized, and an atraumatic clip was used to interrupt the arterial and portal venous blood supply to the cephalad liver lobes for 90 minutes, as described previously.23 The S1PR3 antagonist, CAY10444 (1 mg/kg, IP; Cayman Chemical, Ann Arbor, MI), was administered 30 minutes prior to ischemia.
Hu Y., Yang C., Shen G., Yang S., Cheng X., Cheng F., Rao J, & Wang X. (2019). Hyperglycemia‐Triggered Sphingosine‐1‐Phosphate and Sphingosine‐1‐Phosphate Receptor 3 Signaling Worsens Liver Ischemia/Reperfusion Injury by Regulating M1/M2 Polarization. Liver Transplantation, 25(7), 1074-1090.
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5

Molecular Profiling of Immune Cells

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1640 was from Invitrogen (Grand Island, NY). Fetal bovine serum was from Biochrom (Berlin, Germany). PCR reagents were from Applied Biosystems (Foster City, CA). S1P and dihydro-S1P (H2S1P) were from Biomol (Tebu, France). JTE-013 and CAY-10444 were from Cayman Chemical (Ann Arbor, MI). Anti-F4/80 antibody used for flow cytometry analysis was from BD Biosciences (San Jose, CA). Histopaque-1077 and other common reagents were from Sigma (St. Louis, MO).
Yang L., Han Z., Tian L., Mai P., Zhang Y., Wang L, & Li L. (2015). Sphingosine 1-Phosphate Receptor 2 and 3 Mediate Bone Marrow-Derived Monocyte/Macrophage Motility in Cholestatic Liver Injury in Mice. Scientific Reports, 5, 13423.
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6

Signaling Pathway Antibody Analysis

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Anti-ICAM-1, anti-GAPDH, anti-S1PR1, anti-S1PR2, anti-S1PR3, anti-c-Src, anti-EGFR, anti-PDGFR, anti-JNK1, anti-p42, anti-p38, anti-c-Jun, and anti-c-Fos antibodies and ICAM-1 neutralizing antibody were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-phospho-c-Src, anti-phospho-EGFR, anti-phospho-PDGFR, anti-phospho-JNK1/2, anti-phospho-p42/p44 MAPK, anti-phospho-p38 MAPK, anti-phospho-Akt, and anti-phospho-c-Jun antibodies were from Cell Signaling (Danver, MA). W123, JTE-013 and CAY10444 were from Cayman (Ann Arbor, MI). PP1, U0126, SP600125, SB202190, AG1478, AG1296, Genistein, Tanshinone IIA, and LY294002 were from Biomol (Plymouth Meetings, PA). BCECF/AM was from Molecular Probes (Eugene, OR). SDS-PAGE reagents were from MDBio Inc (Taipei, Taiwan). All other reagents were from Sigma (St. Louis, MO).
Lin C.C., Lee I.T., Hsu C.H., Hsu C.K., Chi P.L., Hsiao L.D, & Yang C.M. (2015). Sphingosine-1-Phosphate Mediates ICAM-1-Dependent Monocyte Adhesion through p38 MAPK and p42/p44 MAPK-Dependent Akt Activation. PLoS ONE, 10(3), e0118473.
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7

Signaling Pathway Inhibitors in Cell Assays

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W 123 (10 µM), JTE 013 (1–10 µM), CAY 10444 (10 µM), specific inhibitors of S1PR1, S1PR2, S1PR3 respectively, S1P (0.1–10 µM), and the EGFR inhibitor tryphostin AG-1478 (0.3–3 µM) were all obtained from Cayman Chemical (Ann Arbor, MI, USA). Helenalin (1 µM), inhibitor of NF-κB, pEGFR antibody (p-Tyr-845 SC-23420-R) and total EGFR antibody (SC-03) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). SR 11302 (1 µM), inhibitor of activator protein-1 (AP-1) and SEW 2871 (10 µm), agonist of S1PR1 were obtained from Tocris Bioscience (Bristol, UK). Dichlorodihydrofluorescein diacetate (DCFH-DA) (10 µM) and N-acetyl cysteine (NAC) (1 mM) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Luciferase reporter lysis buffer was obtained from Promega (Madison, WI, USA). GM6001 (25 µM), the broad-spectrum hydroxamic acid inhibitor of matrix metalloproteinases (MMPs) and TAPI-1 (10 µM), inhibitor or MMPs and tumor necrosis factor-α converting enzyme (TACE) were obtained from Calbiochem (La Jolla, CA). Fura-2 AM (10 µM), and pluronic F127 (0.02%) were obtained from Life Technologies (Carlsbad, Ca).
O’Sullivan M.J., Hirota N, & Martin J.G. (2014). Sphingosine 1-Phosphate (S1P) Induced Interleukin-8 (IL-8) Release Is Mediated by S1P Receptor 2 and Nuclear Factor κB in BEAS-2B Cells. PLoS ONE, 9(4), e95566.
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8

Intraperitoneal Injection of CAY10444

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CAY10444 (Cayman chemical, MI, USA) was dissolved in 1:1 mixture of chremophore EL and 100% ethanol, diluted in water, and injected intraperitoneally to mice at 0.1, 0.2, and 0.5 mg/kg at the time of reperfusion. For the tMCAO group, equal volumes of the vehicle were injected.
Gaire B.P., Song M.R, & Choi J.W. (2018). Sphingosine 1-phosphate receptor subtype 3 (S1P3) contributes to brain injury after transient focal cerebral ischemia via modulating microglial activation and their M1 polarization. Journal of Neuroinflammation, 15, 284.
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9

Reconstitution and Preparation of Bioactive Compounds

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1% bovine serum albumin (BSA) solution was used to dissolve endothelin-1 (Millipore Sigma, St. Louis, MO, USA). Methanol (MeOH) and 0.3M NaOH solution were used to reconstitute S1P (Millipore Sigma, St. Louis, MO, USA and Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturer’s recommendations. W146 (Cayman Chemical & Tocris Bioscience, Minneapolis, MN, USA) was reconstituted in MeOH and 0.03M NaOH. CAY10444 and c-PTIO (Cayman Chemical) were dissolved in dimethylformamide and water, respectively. GKT137831 (Cayman Chemical) and GSK2795039 (Millipore Sigma) were both dissolved in ethanol. Apocynin and PEG-Catalase (Millipore Sigma) were dissolved in MeOH and water, respectively. Acetylcholine (Millipore Sigma) was diluted in water. S1P receptor antibodies were purchased from Invitrogen by Thermo Fisher (Thermo Fisher Scientific, Waltham, MA, USA). ROS-ID® NO Detection kit (Enzo Life Sciences, Enzo Biochem, Farmingdale, New York) was dissolved in HEPES buffer at the time of the experiment.
Katunaric B., SenthilKumar G., Schulz M.E., De Oliveira N, & Freed J.K. (2022). Sphingosine-1-Phosphate-Induced Vasodilation in Human Resistance Arterioles During Health and Disease. Hypertension (Dallas, Tex. : 1979), 79(10), 2250-2261.
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10

Inhibition of TGF-β1 Signaling Pathway

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Recombinant human TGF-β1 was purchased from Merck Millipore. TGF-β1 neutralizing antibody and control IgG antibody (Rabbit polyclonal IgG) were obtained from R&D systems. CAY10444, JTE-013, SB203580, sphingosine-1-phosphate (S1P), SKI-5C (CAY10621), SIS3, and W146 were obtained from Cayman Chemicals. A83-01 was purchased from Sigma-Aldrich. ALK5 inhibitor II was obtained from Enzo Life Sciences.
Beach J.A., Aspuria P.J., Cheon D.J., Lawrenson K., Agadjanian H., Walsh C.S., Karlan B.Y, & Orsulic S. (2015). Sphingosine kinase 1 is required for TGF-β mediated fibroblast-to-myofibroblast differentiation in ovarian cancer. Oncotarget, 7(4), 4167-4182.
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