Cells were cultured in the presence or absence of 10 ng/ml IL-1β (R&D Systems, Inc.) for 24 h. Subsequently, the cells were treated with or without JTE907 (0.6 μM; Tocris Bioscience) for 30 min prior to the addition of WIN-55 (1.0 or 8.0 μM; Tocris Bioscience) for a further 24-h incubation. Transcription inhibitor actinomycin D (1 mg/ml; Sigma-Aldrich) was added. The mRNA level of syndecan-1 was determined with RT-qPCR after 1, 2 and 4 h of actinomycin D treatment, and expressed as fold changes to mRNA level in control cells immediately prior to actinomycin D treatment (designated as 1).
Jte907
Manufactured by Bio-Techne
Sourced in United Kingdom
JTE907 is a small molecule inhibitor developed by Bio-Techne. It functions as a selective inhibitor of the JAK-STAT signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Win 55, by Bio-Techne (2 mentions)
Il 1β, by R&D Systems (2 mentions)
Actinomycin d, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Y 27632, by Merck Group (1 mentions)
Cp55940, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Suramin, by Merck Group (1 mentions)
Oleic acid, by Fujifilm (1 mentions)
Mouse tnf α elisa ready set go, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Mrs2365, by Bio-Techne (1 mentions)
Recombinant tnf α, by Thermo Fisher Scientific (1 mentions)
Fura 2 am, by Merck Group (1 mentions)
Collagenase type xi, by Merck Group (1 mentions)
Guinea pig anti insulin, by Agilent Technologies (1 mentions)
Rabbit anti ki67 primary antibody, by Abcam (1 mentions)
Caspase glo 3 7, by Promega (1 mentions)
Alexa fluor 594 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
4 protocols using jte907
1
Syndecan-1 mRNA Regulation Assay
2
Modulation of Chondrocyte Signaling
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Primary human OA articular chondrocytes (cat no. 402OAK-05a) and human chondrocyte growth medium (cat no. 411–500) were purchased from Cell Applications, Inc. (San Diego, CA, USA). The cells were cultured in the growth medium supplemented with 5% fetal bovine serum (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 100 U/ml penicillin-streptomycin (Sigma-Aldrich, Beijing, China) in an incubator with a humidified atmosphere of 95% air and 5% CO2 at 37°C. The cells were cultured in the presence or absence of 10 ng/ml IL-1β (R&D Systems, Inc., MN, USA) for 24 h. Subsequently, the cells were treated with or without selective CB1 antagonist MJ15 (1, 10 or 50 μM; cat no. 4063; Tocris Bioscience, Bristol, UK) or selective CB2 antagonist JTE907 (0.1, 0.3, or 0.6 μM; cat no. 2479; Tocris Bioscience) for 30 min prior to the addition of WIN-55 (0.5, 1.0, 2.0, 4.0 or 8.0 μM; cat no. 1038; Tocris Bioscience) for a further 24 h.
3
Microglia Activation and Cytokine Regulation
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Isolated microglia were plated at density of 3 × 104 cells per well on PDL-coated plates. For the assays using inhibitors or antagonists, microglia were pretreated with H-89 (protein kinase A, downstream inhibitor of Gαs; Sigma-Aldrich), UBO-QIC (Gq inhibitor)40 (link), Y-27632 (Rho-associated coiled-coil-forming kinase; downstream inhibitor of Gα12/13; Sigma-Aldrich), and JTE907 (CB2-selective antagonist; TOCRIS, Bristol, UK) for 1 hour; with PTX (Gi inhibitor; Wako, Tokyo, Japan) for 6 hours; or with suramin (P2 receptor antagonist; Sigma-Aldrich) for 30 min. After pretreatment, microglia were treated with OAD (Sigma-Aldrich) overnight, and then stimulated with 5 ng/ml of LPS (Sigma-Aldrich) and 0.5 ng/ml of IFN-γ (R&D Systems, MN, USA) for 12 hours. CP-55940 (Sigma-Aldrich), MRS2365 (TOCRIS), LysoPS (P2Y10 agonist12 (link)), P2Y10-selective agonist41 (link) or OAD analogs, including tOAD (Toronto Research Chemicals, Toronto, Canada), oleic acid (Wako, Osaka, Japan), OEtA (Cayman Chemical, MI, USA), OEA (Cayman Chemical), and PEA (Cayman Chemical), were tested as agonists for each GPCR. The concentration of TNF-α in the culture supernatant was quantified by enzyme-linked immunosorbent assay (ELISA) using Mouse TNF-α ELISA Ready-SET-Go (eBioscience, CA, USA).
4
Isolation and Characterization of Pancreatic Cells
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Culture media and supplements, collagenase type XI, histopaque-1077, DMSO, EDTA, IBMX, carbachol, clonidine, Accutase, Fura-2 AM, tolbutamide, LiCl, exendin-4, forskolin, agarose, bionic buffer, and BSA were obtained from Sigma-Aldrich (Dorset, UK). JTE 907 was from Tocris Bioscience (Abingdon, UK). Rabbit anti-Ki67 primary antibody was from Abcam (Cambridge, UK). cAMP HiRange and IP-one (IP1) assays were from Cisbio (Codolet, France). HEPES, HBSS, and DAPI were from Thermo Fisher Scientific (Paisley, UK). Caspase-Glo 3/7 was from Promega (Southampton, UK). Recombinant TNFα, IFNγ, and IL-1β were from PeproTech EC (London, UK). Guinea pig anti-insulin was purchased from Dako (Cambridge, UK). Alexa Fluor 488- and Alexa Fluor 594-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories (Newmarket, UK).
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