Ginkgetin and GSK1016790A (GSK101) were purchased from Sigma-Aldrich (St. Louis, MO), and FLIPR calcium 6 assay kit was obtained from Molecular Device (Sunnyvale, CA). Human native LDL (nLDL) was purchased from Stemcell Technologies (Vancouver, Canada). We incubated nLDL with 5 µM CuSO4 for 12 h at 37 °C to prepare copper-oxidized LDL (oxLDL)7 (link),14 (link),16 . Fluorescent dye-labeled, DiI-oxLDL, was purchased from Invitrogen (Carlsbad, CA), and MCSF from R&D (Minneapolis, MN). Antibodies for FACS analysis were: TRPV4 (Millipore; Burlington, MA), CD36 (R&D; Minneapolis, MN), TLR4 and TLR6 (Novus Biologicals; Centennial, CO). PE-conjugated secondary antibodies were from R&D (Minneapolis, MN), and PE-conjugated isotype controls were from BD (Franklin Lake, NJ). For quantitative polymerase chain reaction (qRT-PCR), we used RNeasy kit from QIAGEN (Redwood City, CA). All primers were purchased from Bio-Rad (Hercules, CA). Cell culture media, cell culture related products, and western blot reagents were obtained from Thermo Fischer Scientific (Waltham, MA).
Gsk1016790a gsk101
Manufactured by Merck Group
Sourced in United States
GSK1016790A (GSK101) is a synthetic compound developed by Merck Group for use in laboratory research. It functions as an agonist for the transient receptor potential cation channel subfamily V member 4 (TRPV4). This channel plays a role in various physiological processes. The specific details and intended use of GSK101 are not provided in order to maintain an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Flipr calcium 6 assay kit, by Molecular Devices (2 mentions)
Hc 067047 hc 06, by Merck Group (2 mentions)
Western blot reagent, by Thermo Fisher Scientific (1 mentions)
Cell culture media, by Thermo Fisher Scientific (1 mentions)
Ginkgetin, by Merck Group (1 mentions)
Dii oxldl, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
M csf, by R&D Systems (1 mentions)
Probenecid, by Cayman Chemical (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Endothelin 1, by Merck Group (1 mentions)
Histamine, by Merck Group (1 mentions)
Compound 48 80, by Merck Group (1 mentions)
U0126, by Selleck Chemicals (1 mentions)
Goat antirabbit alexaflour 488, by Thermo Fisher Scientific (1 mentions)
Anti trpv4, by Alomone (1 mentions)
Anti cd36, by BD (1 mentions)
1 1 dioctadecyl 3 3 3 3 tetramethylindocarbocyanine perchlorate dii, by Merck Group (1 mentions)
Rpmi 1640 cell culture media, by Thermo Fisher Scientific (1 mentions)
Anti β actin hrp conjugate, by Santa Cruz Biotechnology (1 mentions)
11 protocols using gsk1016790a gsk101
1
Copper-Oxidized LDL Preparation and Analysis
2
Pharmacological Modulation of TRPV4 Channel
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The TRPV4 agonist GSK1016790A (GSK101) and antagonist HC-067047 (HC-06) were purchased from Sigma (St. Louis, MO, USA). Suramin and probenecid were obtained from Cayman Chemicals (Ann Arbor, MI, USA). Other salts and reagents were purchased from Sigma, VWR (West Chester, PA, USA), Across Organics (Pittsburgh, PA, USA), or Thermo Fisher (Waltham, MA, USA). GSK101 (1 mM), HC-06 (20 mM), probenecid (50 mM), and suramin (100 mM) stocks were diluted in extracellular saline solution to working concentrations before use. After dissolution in dimethyl sulfoxide (DMSO), the final drug concentrations did not exceed 0.1%.
3
Pharmacological Agents Preparation
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Histamine, compound 48/80, endothelin-1, chloroquine, and GSK1016790A (GSK101) were purchased from Sigma. GSK2193874 (GSK219) was obtained from Tocris, U0126 was from Selleckchem, and GSK205 was synthesized (26 (link), 33 (link)). All were dissolved in sterile normal saline except that GSK101 and GSK205 were dissolved in DMSO (20 mm in stock) and further diluted until use.
4
Molecular Mechanisms of Macrophage Activation
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TRPV4‐specific antagonist, GSK2193874 (GSK219), TRPV4‐specific agonist, GSK1016790A (GSK101), nucleus‐specific stain, DiI (1,1′‐Dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate), and Ca2+ ionophore, A23187 (A23) were purchased from Sigma‐Aldrich (St. Louis, MO). The following antibodies were purchased: anti‐TRPV4 (Alomone Labs; Jerusalem, Israel); anti‐β‐Actin HRP‐conjugate (Santa Cruz Biotechnology; Dallas, TX); anti‐CD36 (BD Pharmingen); goat‐anti rabbit Alexa Flour 488 and goat‐anti Mouse Alexa Flour 594 (Life technologies). FLIPR Calcium 6 assay kit was purchased from Molecular Devices (Sunnyvale, CA) and macrophage colony‐stimulating factor (M‐CSF) was obtained from R & D. We prepared copper‐oxidized LDL (oxLDL) by incubating human native LDL (Stemcell Technologies; Vancouver, BC, Canada) with CuSO4 (5 μm) for 12 h at 37°C, as described previously (Rahaman et al. 2006 , 2013 ). Cell culture media (RPMI‐1640) and other cell culture‐related reagents were purchased from Gibco. We obtained collagen‐coated polyacrylamide hydrogels (0.5‐50 kPa) from Matrigen Life Technologies (Brea, CA). P. gingivalis ‐derived lipopolysaccharide was purchased from InvivoGen (San Diego, CA).
5
Chondrocyte Osmotic Response Imaging
6
Compounds for Skin Inflammation Models
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Compounds flopropione (MW: 182.18) and osthole (MW: 244.29) were purchased from TargetMol. Compounds 2-aminoethoxydiphenyl borate (2-APB), carvacrol (Car), capsaicin, menthol, GSK1016790 A (GSK101), and allyl isothiocyanate (AITC) were purchased from Sigma-Aldrich. Compounds were made in DMSO as stock solutions before use. Compounds used for patch-clamp recordings were diluted in perfusion solution. Compounds used for generation of atopic dermatitis and ear swelling models were diluted in the solvent of acetone/olive oil = 4/1, v/v before topical applications.
7
Investigating the NF-κB Pathway
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For investigation of the NF-κB pathway, IκBα, p65, and phospho-p65 antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Vimentin, nestin, and GAPDH antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The Human Quantikine TNF-α ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA). The GFAP antibody and RIPA Cell Lysis buffer were purchased from ThermoFisher Scientific (Fair Lawn, NJ, USA). GSK1016790A (GSK101) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
8
Pharmacological Modulation of TRPV1/TRPV4 in RGCs
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The TRPV4 agonist GSK1016790A (GSK101) and antagonist HC-067047 (HC-06) were purchased from Sigma. The TRPV1 agonist capsaicin (CAP; 8-methyl-N-vanillyl-6-nonenamide) and the TRPV1 antagonist capsazepine (CPZ; N-[2-(4-Chlorophenyl)ethyl]-1,3,4,5-tetrahydro-7,8-dihydroxy-2H-2-benzazepine-2-carbothioamide) and the endogenous agonist of CB1 receptors 2-arachidonoylglycerol (2-AG) were obtained from Cayman Chemicals. BDNF and CNTF used to culture RGCs were obtained from GenWay Biotech. Other salts and reagents were purchased from Sigma, VWR, Across Organics, or Thermo Fisher. GSK101 (1 mM), HC-06 (10 mM), CAP (10 mM), and CPZ (20 mM) stocks in DMSO were diluted in extracellular saline before use and placed into reservoirs connected to gravity-fed perfusion systems (Warner Instruments).
9
TRPV4 Agonist GSK101 Protocol
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The TRPV4 agonist GSK1016790A (GSK101) and cholesterol were obtained from Sigma or VWR. GSK101 (1 mM) stock aliquots were prepared in dimethyl sulfoxide and subsequently diluted into working saline concentrations (5 and 25 nM, respectively). Chemical reagents for biochemical experiments—methanol, isopropanol, n-hexane, and chloroform—were of GC/MS grade and purchased from Thermo Fisher Scientific. The cholesterol standard was purchased from Sigma-Aldrich.
10
Calcium Signaling Modulators Protocol
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TRPV4 specific agonist GSK1016790A (GSK101, #G0798) and inhibitor GSK2193874 (GSK874, #5106) were obtained from Sigma-Aldrich (St. Louis, MO) and Tocris Bioscience (Bristol, UK), respectively. Compound C (AMPK inhibitor, #11967), STO-609 (calmodulin-dependent kinase kinase (CaMKK) inhibitor, #15325), Wortmannin (PI3K inhibitor, #10010591), LY294002 (Akt inhibitor, #70920), U0126 (ERK1/2 inhibitor, #70970), AICAR (#10010241), Forskolin (#11018), and Phorbol 12-myristate 13-acetate (PMA, #10008014) were obtained from Cayman Chemical (Ann Arbor, MI). Ruthenium Red (Ca2+ fluxes blocker, #557450), and H-89 (PKA inhibitor, #371963) were obtained from Calbiochem (San Diego, CA). Calcium chelator BAPTA/AM (#B-1205) was from Invitrogen (Grand Island, NY). Recombinant mouse TNF-α was from Roche (#11271156001, Indianapolis, IN). Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME, #N5751), Sodium nitroprusside (SNP, #PHR1423), Acetylcholine chloride (#A2661), (R)-(−)-Phenylephrine hydrochloride (PE, #P8155) and other chemicals were obtained from Sigma-Aldrich.
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