Gkt137831

Manufactured by MedChemExpress

Sourced in United States, Sweden

GKT137831 is a chemical compound that functions as a selective inhibitor of NOX4, a member of the NADPH oxidase (NOX) family of enzymes. It is utilized as a research tool to study the role of NOX4 in various biological processes.

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Lab products found in correlation

4 protocols using gkt137831

Evaluating Apoptosis-Inducing Pathways

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Without specific indication, the reagents purchased from Sigma‐Aldrich Inc. were used in the present study, and cell culture supplements were from GIBCO/Life Technologies Inc. Vincristine (VCR), SZLP1‐41 (Skp2 inhibitor), E64D (lysosomal protease inhibitor) and epoxomicin (proteasome inhibitor) were the products of Apexbio Technology LLC. GKT137831 and GLX351322 were purchased from MedChem Express; MitoSOX, annexin V‐FITC/propidium iodide (PI) apoptosis detection kit, tetramethylrhodamine (TMRM) and H₂DCFDA were from Molecular Probes (Eugene, OR). Antibodies against caspase‐3 and caspase‐8, okadaic acid, Z‐IETD‐FMK and Z‐DEVD‐FMK were purchased from Calbiochem. Antibodies separately against PP2Ac, Mcl‐1, tristetraprolin (TTP), Fas, FasL, TNF‐α and FADD were purchased from Santa Cruz Biotechnology Inc.; and antibody against TNFR1 was from R&D Systems. Antibodies separately against ERK, p‐ERK, p38 MAPK, p‐p38 MAPK, JNK, p‐JNK, SIRT3, TNFR2, α‐tubulin, MID1, α4, caspase‐9, Bax, Bak and Bcl‐2 were obtained from Cell Signaling Technology. Antibodies separately against cytochrome c, Bcl‐xL and Bid were the products of BD Pharmingen Technical (San Jose, CA), and anti‐NOX4 antibody was from Novus Biologicals. Secondary antibodies conjugated with horseradish peroxidase (HRP) were the products of Pierce (Rockford, IL).

Wang L., Chiou J., Lee Y., Huang C., Shi Y, & Chang L. (2020). SIRT3, PP2A and TTP protein stability in the presence of TNF‐α on vincristine‐induced apoptosis of leukaemia cells. Journal of Cellular and Molecular Medicine, 24(4), 2552-2565.

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Mitochondrial Dysfunction in Cigarette Smoke-Induced Lung Injury

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PM2.5 (SRM1649b) was obtained from NIST (MD, USA). Cigarettes were purchased from Hongta Tobacco (Group) Co., Ltd. (Yunnan, China). GKT137831 and mito-TEMPO were obtained from Med Chem Express (New Jersey, USA). Specific antibodies against Nrf2 (ab137550), p-mTOR (ab109268), SIRT3 (ab189860), VCAM (ab174279), ICAM (ab179707) and β-actin (ab8226) were purchased from Abcam (Cambridge, UK). Specific antibodies against caspase3 (9662S), Bax (5023S), Bcl2 (3498S), cleaved caspase-3 (9664S), Drp1 (8570S), AIF (4642), Cyto-c (4272), LC3B (2775S), P62 (5114S), ATG3 (3415), Tom20 (42406S), and Parkin (4211S) were purchased from Cell Signaling Technology (Danvers, USA). A specific antibody against PINK1 (P0076) was provided by Sigma–Aldrich (St. Louis, MO, USA). Specific antibody against NOX4 (Cat NO.380874) and MUC5ac (Cat NO.381811) were obtained from ZENBIO (Nanjing, China). Specific antibodies against Tim23 (Cat No.11123-1-AP) and p-creb (Cat No.28792-1-AP) were obtained from Proteintech (Chicago, USA). The BCA protein assay kit (lot # tk274307, USA) was obtained from Thermo Fisher Scientific. NOX4 siRNA was obtained from HanBio Technology (Shanghai, China). Cell Counting Kit-8 (CCK-8) and LDH kits were obtained from Beyotime (Shanghai, China).

Fan X., Dong T., Yan K., Ci X, & Peng L. (2022). PM2.5 increases susceptibility to acute exacerbation of COPD via NOX4/Nrf2 redox imbalance-mediated mitophagy. Redox Biology, 59, 102587.

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Evaluating Vascular Function in Docetaxel-Treated Mice

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Studies of mice were conducted according to the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines. We used male C57BL/6J and Nox4^–/– (B6.129-Nox4tm1Kkr/J) mice obtained from The Jackson Laboratory between the ages of 3.5 and 4.5 months. Mice were housed in the animal facility at Jagiellonian University Medical College and at the Faculty of Biochemistry, Biophysics and Biotechnology (Jagiellonian University).
Mice were treated with docetaxel (10 mg/kg, Cayman Chemicals) or placebo (solvent) i.p. every 5 days for 3 weeks. GKT137831 (40 mg/kg, MedChemExpress) or fasudil (30 mg/kg; MedChemExpress) was injected s.c. daily during docetaxel treatment. After this, mice were euthanized using CO₂ inhalation, and left ventricle perfusion was performed using cold PBS. Aortic rings were used to measure vascular function, ROS generation, and gene and protein expression.

Szczepaniak P., Siedlinski M., Hodorowicz-Zaniewska D., Nosalski R., Mikolajczyk T.P., Dobosz A.M., Dikalova A., Dikalov S., Streb J., Gara K., Basta P., Krolczyk J., Sulicka-Grodzicka J., Jozefczuk E., Dziewulska A., Saju B., Laksa I., Chen W., Dormer J., Tomaszewski M., Maffia P., Czesnikiewicz-Guzik M., Crea F., Dobrzyn A., Moslehi J., Grodzicki T., Harrison D.G, & Guzik T.J. (2022). Breast cancer chemotherapy induces vascular dysfunction and hypertension through a NOX4-dependent mechanism. The Journal of Clinical Investigation, 132(13), e149117.

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Quantifying Cellular Reactive Oxygen Levels

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ROS production was monitored by using the 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-fluorescein di-acetate, acetyl ester assay (C6827; Life Technologies, Carlsbad, CA) as previously described. 38 (link) After a 4-hour pretreatment, cells were washed with DPBS and loaded with 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-fluorescein diacetate, acetyl ester (10 μmol/L, 30 minutes, in the dark). Cells were washed thoroughly with DPBS, and fluorescence intensity was measured in every 30 minutes for 4 hours applying 488 nm excitation and 533 nm emission wavelengths. In some experiments, we applied ROS inhibitors during the hypoxia treatment, N-acetyl cysteine (NAC, 1 mmol/L; A9165; Sigma), Mn(III)-tetrakis (4-benzoic acid) porphyrin chloride (50 μmol/L; sc-221954; Santa Cruz Biotechnology, Inc), sodium pyruvate (SP, 5 mmol/L; P5280; Sigma), GKT137831 (GKT, 20 μmol/L; HY-12298; MedChem Express, Sollentuna, Sweden), and rotenone (5 μmol/L; R8875; Sigma).

Balogh E., Tóth A., Méhes G., Trencsényi G., Paragh G, & Jeney V. (2019). Hypoxia Triggers Osteochondrogenic Differentiation of Vascular Smooth Muscle Cells in an HIF-1 (Hypoxia-Inducible Factor 1)-Dependent and Reactive Oxygen Species-Dependent Manner. Arteriosclerosis, thrombosis, and vascular biology, 39(6).

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