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A438079 hydrochloride

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A438079 hydrochloride is a chemical compound used in laboratory research. It is a selective inhibitor of the enzyme cyclin-dependent kinase 9 (CDK9). CDK9 plays a role in the regulation of gene expression and cellular processes. The product is provided as a hydrochloride salt formulation.

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Lab products found in correlation

Atp disodium salt hydrate, by Merck Group (1 mentions) Macs human cd14 microbeads, by Miltenyi Biotec (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions) Ar c 118925xx, by Bio-Techne (1 mentions) Cellmask deep red plasma membrane stain, by Thermo Fisher Scientific (1 mentions) Vectashield with dapi, by Vector Laboratories (1 mentions) 2 3 o 4 benzoylbenzoyl atp bzatp, by Merck Group (1 mentions) Fluo 3am fluorescent dye, by Thermo Fisher Scientific (1 mentions) 10panx inhibitory peptide, by Bio-Techne (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Necrosulfonamide nsa, by Bio-Techne (1 mentions) Adenosine triphosphate (atp), by InvivoGen (1 mentions) Brefeldin a, by BioLegend (1 mentions) Monensin, by BioLegend (1 mentions) Legato 200 syringe pump, by KD Scientific (1 mentions) μ slide vi0.4 chamber, by Ibidi (1 mentions) A740003, by Bio-Techne (1 mentions) Methamphetamine hydrochloride, by Merck Group (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)

7 protocols using a438079 hydrochloride

1

ATP-Induced Monocyte Transcriptional Regulation

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Monocytes from individuals heterozygous for rs257174 were purified from PBMCs by positive selection using MACS human CD14 MicroBeads (Miltenyi Biotec) according to manufacturer’s instructions. Monocytes were plated at 0.5 × 106 cells/well in 24-well tissue culture plates and then incubated with or without cell culture-grade ATP disodium salt hydrate (Sigma-Aldrich) in RPMI-1640 supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 units/ml of penicillin and 100 μg/ml of streptomycin. For the inhibition of ATP the incubation was carried out in the presence of 10 μM of the purinergic receptor inhibitor A-438079 hydrochloride (Tocris Bioscience). Monocytes were incubated for 2 h before harvesting using cell scrapers. The treated monocytes were stored in TRIzol (Invitrogen Life Technologies) at -80°C until further analysis.
Melchiotti R., Puan K.J., Andiappan A.K., Poh T.Y., Starke M., Zhuang L., Petsch K., Lai T.S., Chew F.T., Larbi A., Wang D.Y., Poidinger M, & Rotzschke O. (2014). Genetic analysis of an allergic rhinitis cohort reveals an intercellular epistasis between FAM134B and CD39. BMC Medical Genetics, 15, 73.
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2

Molecular Probes for Cell Signaling

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ATP, 2,3-O-(4-benzoylbenzoyl)-ATP (BzATP), and UTP were all purchased from Sigma-Aldrich (St. Louis, MO). A luciferase-based ATP Determination Kit was purchased from Invitrogen (Carlsbad, CA). A438079 hydrochloride, AR-C 118925XX, 10Panx Inhibitory peptide, and scramble control peptide were purchased from Tocris Biosciences (Minneapolis, MN). Pannexin-1 polyclonal rabbit antibodies were purchased from Alomone (Jerusalem, Israel), and Connexin-43 (Cx43) polyclonal rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Fluo-3AM fluorescent dye, Rhodamine-phalloidin and secondary antibodies (Alexa Fluor-conjugated secondary antibody) were purchased from Invitrogen (Carlsbad, CA). CellMask Deep Red Plasma membrane stain was purchased from ThermoFisher (Waltham, MA) and SiR-Actin Spirochrome probe was purchased from Cytoskeleton Inc. (Denver, CO). VectaSHIELD with DAPI was purchased from Vector Labs (Burlingame, CA).
Lee Y., Kim M.T., Rhodes G., Sack K., Son S.J., Rich C.B., Kolachalama V.B., Gabel C.V, & Trinkaus-Randall V. (2019). Sustained Ca2+ mobilizations: A quantitative approach to predict their importance in cell-cell communication and wound healing. PLoS ONE, 14(4), e0213422.
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3

Modulation of ATP-induced Inflammation

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LPS-stimulated IC21 cells were co-stimulated for 1h with 3mM ATP (In vivoGen, CA), in the presence or absence of 2 µM monensin (Biolegend, CA), 5 µg/ml brefeldin A (Biolegend, CA), 200nM A438079 hydrochloride (TOCRIS, UK) or 20µM Necrosulfonamide (NSA; TOCRIS, UK) (specific inhibitors specific for P2X7R and Gasdermin D respectively). Supernatants and protein lysates were evaluated after co-stimulation.
Manzanares-Meza L.D., Gutiérrez-Román C.I., Jiménez-Pineda A., Castro-Martínez F., Patiño-López G., Rodríguez-Arellano E., Valle-Rios R., Ortíz-Navarrete V.F, & Medina-Contreras O. (2022). IL-36γ is secreted through an unconventional pathway using the Gasdermin D and P2X7R membrane pores. Frontiers in Immunology, 13, 979749.
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4

Oscillatory Fluid Shear Stress Responses

Cited 3 times
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Oscillatory fluid shear stress (OFSS, τ = ± 10 dyne/cm2) at 1 Hz was applied over a cell monolayer. The fluid flow setup consisted of μ–slide VI0.4 chamber (ibidi GmbH, Germany), Legato 200 syringe pump (KD Scientific, MA, USA) and a syringe filled with flow media (phenol free α-MEM supplemented with 1% FBS and 10mM Hepes) that is optimized for real-time imaging. Culture media in μ–slide VI0.4 chambers was first replaced with flow media followed by flow-loop set up and equilibrated under static condition for 2 min prior to OFSS. Aliquot (50 μl) of static media was collected right after 2 min equilibration time and flow conditioned media was collected at the end of 5 min OFSS exposure for measurements of cellular ATP release.
To inhibit P2X7R and Panx1 function, cells were pretreated with the P2X7R blockers A438079 hydrochloride (10 μM; TOCRIS bioscience, Bristol, United Kingdom) [41 (link), 51 (link)], A740003 (10 μM; TOCRIS) [30 (link), 52 (link)] and mefloquine (MFQ, 100nM; QU024-1, Bioblocks, San Diego, CA) [26 (link)] in flow media for 15 min prior to OFSS and during 5 min of OFSS.
Seref-Ferlengez Z., Maung S., Schaffler M.B., Spray D.C., Suadicani S.O, & Thi M.M. (2016). P2X7R-Panx1 Complex Impairs Bone Mechanosignaling under High Glucose Levels Associated with Type-1 Diabetes. PLoS ONE, 11(5), e0155107.
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5

Methamphetamine Addiction Model in Mice

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Adult male C57Bl/6 mice (Jackson Laboratories; Bar Harbor, ME) were group-housed (2–5/cage) and maintained under a 12-h light/dark cycle on a regular chow diet. Studies were conducted in accordance with the Institutional Animal Care and Use Committee at Temple University. Methamphetamine hydrochloride (METH) was purchased from Sigma-Aldrich (St. Louis, MO). A438079 hydrochloride was purchased from Tocris Biosciences (St. Louis, MO, USA). Drugs were dissolved in 0.9% saline and injected intraperitoneally (ip). Mice were assigned randomly to each experimental group and used only once. Testing was done by an experimenter blinded to treatment conditions.
Tsiokas L., Arnould T., Zhu C., Kim E., Walz G, & Sukhatme V.P. (1999). Specific association of the gene product of PKD2 with the TRPC1 channel. Proceedings of the National Academy of Sciences of the United States of America, 96(7), 3934-3939.
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6

MDPV and A438079 in C57BL/6 Mice

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Adult male C57B1/6 mice (Jackson Laboratories; Bar Harbor, ME) were group-housed (2-5/cage), kept in a humidity- and temperature-controlled vivarium, and maintained under a 12-h light/dark cycle on a regular chow diet. Studies were conducted in accordance with the Institutional Animal Care and Use Committee at Temple University. (±)-MDPV was synthesized according to published methods (Doylestown, PA, USA) (Gregg et al., 2015 (link)). A438079 hydrochloride was purchased from Tocris Biosciences (St. Louis, MO, USA). Drugs were dissolved in 0.9% saline and administered in a volume of 1 ml/kg subcutaneously (SC) (MDPV) or injected intraperitoneally (IP) (A438079).
Gentile T.A., Simmons S.J., Tallarida C.S., Su S., Rom S., Watson M.N., Reitz A.B., Potula R, & Rawls S.M. (2019). Synthetic cathinone MDPV enhances reward function through purinergic P2X7 receptor-dependent pathway and increases P2X7 gene expression in nucleus accumbens. Drug and alcohol dependence, 197, 22-27.
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7

Monocyte Activation by LPS and P2X7R Modulation

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Primary blood monocytes were cultured in RPMI-1640 medium supplemented with 10% (v/v) heat inactivated, low endotoxin fetal calf serum (HI FCS-LE, Gibco), 1% (v/v) Penicillin-Streptomycin (P/S) and 1% (v/v) L-glutamine. Subsets of monocytes at 12,500 cells per well (50,000 cells/ml) were plated in 96 well plates and stimulated with 1 ng/ml LPS (purified from E. coli, serotype 0111:B4, TLRgrade, Enzo Life Sciences) for 24 h either in the presence or absence of 10 μM A438079 hydrochloride (P7X7R antagonist, Tocris Bioscience). 300 μM BzATP (P2X7R agonist, Sigma-Aldrich) was added for the final 20 minutes of incubation. Culture supernatants were collected and non-adherent cells removed by centrifugation. Samples were stored at −80 °C until analysis.
Hadadi E., Zhang B., Baidžajevas K., Yusof N., Puan K.J., Ong S.M., Yeap W.H., Rotzschke O., Kiss-Toth E., Wilson H, & Wong S.C. (2016). Differential IL-1β secretion by monocyte subsets is regulated by Hsp27 through modulating mRNA stability. Scientific Reports, 6, 39035.
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