γ‐secretase inhibitor MRK‐003 was a gift of Merck & Co. (Whitehouse Station, NJ). 5 μM MRK‐003 was used in most experiments; exceptions are reported in figures and/or legends. Torin 1 (1‐[4‐[4‐(1‐Oxopropyl)‐1‐piperazinyl]‐3‐(trifluoromethyl) phenyl]‐9‐(3‐quinolinyl)‐benzo[h]‐1,6‐naphthyridin‐2(1H)‐one) (Tocris Bioscience, Bristol, UK) was dissolved in DMSO and used at a final concentration of 0.250 μM. Roscovitine (Cell Signaling) was dissolved in DMSO and used at a final concentration of 20 μM. UO126 (Selleck Chemicals, Huston, TX) was dissolved in DMSO, and used at a final concentration of 10 μM. Silvestrol (Medchemexpress LLC, Princeton, NJ) was dissolved in DMSO and used at a final concentration of 0.04 μM and 0.1 μM. Homoharringtonine (Tocris) was dissolved in DMSO and used at a final concentration of 0.1 μM. Insulin in solution (Sigma–Aldrich) was diluted in sterile PBS and used at the specified concentrations. pcDNA3‐RLUC‐POLIRES‐FLUC was a gift from Nahum Sonenberg (McGill University, Montreal); pCDF1‐MCS1‐EF1‐cop GFP expressing the APP C‐terminal 59 aa was a gift from Dr. Xiao Z.C. (Institute of Molecular and Cell Biology, Singapore); full length APP 695 in pCAX vector was from Addgene (Cambridge, MA). We used the empty plasmids as transfection controls.
Uo126
Manufactured by Selleck Chemicals
Sourced in United States
UO126 is a chemical compound used in laboratory research. It functions as a selective inhibitor of the mitogen-activated protein kinase (MAPK) pathway, specifically targeting the extracellular signal-regulated kinase (ERK) activation. The core function of UO126 is to inhibit the phosphorylation of ERK1 and ERK2, key components of the MAPK signaling cascade.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Roscovitine, by Cell Signaling Technology (2 mentions)
Silvestrol, by MedChemExpress (2 mentions)
Homoharringtonine, by Bio-Techne (2 mentions)
Insulin in solution, by Merck Group (2 mentions)
Full length app 695 in pcax vector, by Addgene (2 mentions)
Mg132, by Selleck Chemicals (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Torin1, by Bio-Techne (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Apocynin, by Selleck Chemicals (1 mentions)
Telmisartan, by Merck Group (1 mentions)
Diphenyleneiodonium dpi, by Merck Group (1 mentions)
Uric acid, by Merck Group (1 mentions)
Bq788, by Merck Group (1 mentions)
Bq123, by Selleck Chemicals (1 mentions)
Pd98059, by Selleck Chemicals (1 mentions)
Gdc 0973, by Selleck Chemicals (1 mentions)
Plx4032, by Selleck Chemicals (1 mentions)
13 protocols using uo126
1
Investigating Gamma-Secretase Inhibitors
2
Breast Cancer Cell Line Characterization
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Human breast cancer and normal cell lines used in this study were purchased from the Cell Biology Institute of Shanghai, Chinese Academy of Science. The TNBC cell lines MDA-MB-468, MDA-MB-231, and SUM149, and the non-TNBC cell lines MCF-10A, MCF-7, and SKBR3 were used. Cells were cultured as described previously (8, 9) in 5% CO2 saturated humidity at 37 °C. The phosphorylated AKT and phosphorylated ERK inhibitors, MK2206 and UO126, respectively, were purchased from Selleckchem (Houston, TX, USA).
3
Investigating Endothelial Cell Signaling
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Dulbecco's Modified Eagle medium (DMEM) and penicillin and streptomycin were purchased from HyClone (Logan, UT, USA). 10% fetal bovine serum (FBS) was obtained from Sijiqing Biological Engineering Materials Co. (Hangzhou, China). Uric acid, telmisartan, diphenylene iodonium (DPI), and ET(B) receptor antagonist BQ788 were purchased from Sigma (St. Louis, MO, USA). Apocynin, PD98059, UO126, and ET(A) receptor antagonist BQ123 were purchased from Selleck Chemicals (Houston, TX, USA).
4
Modulation of Cellular Signaling Cascades
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I-BET151 was supplied by GlaxoSmithKline (Brentford, UK). BMS-345541, BKM120, BEZ235, GSK1120212-(Trametinib), GDC0973-(Cobemitinib) UO126, PLX4032 (Vemurafenib), GSK2118436 (Dabrafenib), LY294002, were purchased from Selleck Chemicals (Houston, TX, USA). IFN-γ was from Shenandoah Biotechnologies (Warwick, PA, USA).
5
Inhibition of MAPK Pathways in Lung Cells
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The bronchial epithelial cell line BEAS2B, an immortalized cell line transformed using an adenovirus 12–SV40 viral vector, was purchased from Bogoo Biotechnology (Shanghai, China) and cultured in Roswell Park Memorial Institute (RPMI) 1640 complete culture medium containing 10% fetal bovine serum (FBS). Healthy 4- to 6-week-old Sprague Dawley rats of specific pathogen-free (SPF) grade with body weights of 200±20 g were purchased from the Department of Laboratory Animal Science of Fudan University and housed in an SPF-grade experimental animal center at Fudan University. The experimental protocol was approved by the ethics committee of Fudan University and followed the Guide for the Care and Use of Laboratory Animals. UO126 (ERK inhibitor), SP600125 (JNK inhibitor), and SB203580 (p38 inhibitor) were purchased from Selleck (S1102, S1460, and S1076; Shanghai, China). The concentration used in cell experiments was 20 μM, in accordance with a previous report,10 (link) and dosages used in animal experiments were 1 mg/kg, 1.5 mg/kg, and 1 mg/kg, respectively.
6
Cultivation of Human Pancreatic Cancer Cell Lines
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The human pancreatic cancer cell lines SW1990, PANC-1, CFPAC-1, Capan-1, MIA PaCa-2 and AsPC-1 with KRAS mutations were obtained from the American Type Culture Collection. The SW1990 cells were cultured in L-15 medium supplemented with 10% fetal bovine serum (FBS). PANC-1 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS. CFPAC-1 cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) supplemented with 10% FBS. Capan-1 cells were cultured in IMDM supplemented with 20% FBS. MIA PaCa-2 cells were cultured in DMEM supplemented with 10% FBS and 2.5% horse serum. AsPC-1 cells were cultured in Roswell Park Memorial Institute medium supplemented with 10% FBS. The HPDE cells were cultured in complete keratinocyte serum-free medium supplemented with 50 μg/mL bovine pituitary extract and 5 ng/mL epidermal growth factor (Gibco). All of the cell culture media contained 100 U/mL penicillin and 100 mg/mL streptomycin. UO126, AZD6244, trametinib, SCH772984 and MG132 were purchased from Selleck. CHX, CIP and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich.
7
Cytotoxic Treatments for Renal Cell Carcinoma
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RCC cell lines and HEK293T cells were grown at 37 °C in an atmosphere containing 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% foetal bovine serum (FCS), 2 mM glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin. Transduced RCC cells were maintained in the medium described above supplemented with 2 μg/ml puromycin. For treatment, cells were exposed to the following substances dissolved in culture medium for defined periods of time: 0.1 to 10 μg/ml topotecan (Hycamtin®, GSK, Buehl, Germany), 100 ng/ml superkillerTrail (EnzoLifeScience, Lörrach, Germany) or 10 or 20 μM ABT263 (Navitoclax®, Selleckchem, Texas, USA) or 50 μM UO126 (Selleckchem). The corresponding negative controls were prepared with the appropriate solvent (PBS or DMSO).
8
Evaluating Cancer Cell Signaling Pathways
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The cisplatin modified DNA (ab103261) and γH2A.X (ab26350) antibodies were from Abcam (Cambridge, USA). The phospho-STAT3 S727 (#9134), phospho-Akt S473 (#9271), phospho-NF-κB S536 (#3033), pERK T202/Y204 (#9101), pATF2 T71 (#5112), (P70S6K (#2708), phospho-P70S6K T389 (#9205), p21 (#2947), cleaved caspase 3 (#9661) and cleaved caspase 7 (#9491) antibodies were from Cell Signaling (MA, United States). The γH2A.X (S139) antibody (AB26350) was from Abcam (MA, USA). The p53 antibody (sc-126) was from Santa Cruz Biotechnology (TX, USA). The p63 antibody (NB100-691) was from Novus Biologicals (CO, USA). The actin monoclonal antibody (AC-15) was from Sigma-Aldrich (MO, United States). The SignalSilence p70/85 S6 Kinase siRNA was from Cell Signaling (MA, United States). The plasmids for FUCCI live cell imaging, mVenus-hGeminin(1/110) and mCherry-hCdt1(30/120), were a kind gift from Dr Atsushi Miyawaki (Riken, Japan). Dactolisib (NZP-BEZ235), MK2206, S3I-201, UO126 and SC75741 were all from Selleck Chem (MA, USA).
9
Investigating Gamma-Secretase Inhibition
10
Coral Nubbin Incubation with UO126
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Incubations were performed in 100 mL beakers containing one coral nubbin each, and filled with 40 mL of 0.45 μm filtered seawater. They were placed in the dark for one hour in either a control condition containing 0.005% DMSO (vehicle) or a condition with 5 μmol.L
-1 UO126 (Selleck Chemicals), a MEK inhibitor (
Tang
et al., 2003 ). The incubation medium was continuously stirred using magnetic stirrers at a constant temperature of 25°C. At the end of the incubation, nubbins were frozen and kept at – 80°C prior to western blot analysis.
-1 UO126 (Selleck Chemicals), a MEK inhibitor (
et al., 2003
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