Pd98059

Manufactured by Thermo Fisher Scientific

Sourced in United States

PD98059 is a chemical compound that inhibits the activation of the Mitogen-Activated Protein Kinase (MAPK) signaling pathway by selectively inhibiting the enzyme MEK1/2. This compound is commonly used in cell biology research to study the role of the MAPK pathway in various cellular processes.

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Lab products found in correlation

29 protocols using pd98059

Th17 Cell Differentiation Protocol

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The cell culture plate was coated with 2 μg/mL anti-human CD3 antibody (Catalogue No. 05121-25-500, BioGems, Rocky Hill, CT, USA) overnight at 4°C. After washing with PBS, isolated CD4+ T cells were suspended at a density of 1 × 10⁶ cells/mL and seeded with culture medium of X-VIVO 15 serum-free medium (Catalogue No. 04-418Q, Lonza, Alpharetta, GA, USA) containing 1 μg/mL anti-human CD28 antibody (Catalogue No. 10311-25-100, BioGems, Rocky Hill, CT, USA) and Th17 cell differentiation-related cocktail: IL-6 (30 ng/mL), TGF-β (2.25 ng/mL), IL-23 (40 ng/mL), IL-1β (30 ng/mL), anti-IFN-γ (5 μg/mL) and anti-IL-4 (5 μg/mL) (Catalogue Nos. 96-200-06-5, 96-200-23-2, 96-100-21C-2, 96-200-01B-2, 80812-25-100, and 81121-25-500; PeproTech, Rocky Hill, CT, USA).
The inhibition of MEK was performed by adding 10 μM of PD98059, which blocks the phosphorylation and activation of ERK1/2 with no effect on CD4+ T-cell proliferation.[18 (link)] DMSO was used as a reagent control for PD98059. The control group was not added to a cytokine cocktail. Cells were cultured for 4 days and subsequently analysed by flow cytometry, qPCR and ELISA.

Han K., Li S.S., Pan W., Xu M.N., Zhong M.Z., Zhang W.J., Huang X.W, & Zeng K. (2024). ERK/MEK Pathway Regulates Th17 Cell Differentiation in Patients with Pemphigus Vulgaris. Indian Journal of Dermatology, 68(6), 724.

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Lysophosphatidic Acid Signaling Mechanisms

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Recombinant human ATX (also known as Ectonucleotide Pyrophosphatase/Phosphodiesterase-2) was purchased from R&D Systems, Inc., Minneapolis, MN, USA. 1-Oleoyl lysophosphatidic acid sodium salt was from Tocris Bioscience (Bristol, UK). 17-β estradiol (estrogen) was from Sigma-Aldrich; Merck KGaA (Darmstadt, Germany). PD98059 [the phosphorylated extracellular signal-regulated kinase (p-ERK) inhibitor] was from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). ATX antibody for western blotting was from Abcam (Cambridge, UK; ab77104) and for immunohistochemistry from Santa Cruz Biotechnology (Texas, USA; sc-374222). Antibodies against LPA receptors (LPA1, LPA2, LPA3) were from Abcam (ab166903, ab38322 and ab219267, respectively). Total/phosphorylated extracellular signal-regulated kinases (t/p-ERK) were from Cell Signaling Technology, Inc. Danvers, MA, USA (4695s and 4370s). Antibodies against GAPDH were from Santa Cruz Biotechnology (sc-51907). Crystal violet staining solution was from Tiangen Biotech Co., Ltd. (Beijing, China). Cell Counting kit (CCK)-8 was from Beyotime Institute of Biotechnology, Haimen, China (C0037). Transwell culture plates were from Corning Incorporated (Corning, NY, USA). Lipofectamine 2000™ transfection reagent was from Invitrogen, Thermo Fisher Scientific, Inc.

Zhang G., Cheng Y., Zhang Q., Li X., Zhou J., Wang J, & Wei L. (2018). ATX-LPA axis facilitates estrogen-induced endometrial cancer cell proliferation via MAPK/ERK signaling pathway. Molecular Medicine Reports, 17(3), 4245-4252.

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Cytotoxic Agents and Inhibitors

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5-Fluorouracil was purchased from Sigma (Sigma-Aldrich Corporation, Los Angeles, CA, USA), and MAPK/ERK inhibitor (PD98059) and p53 inhibitor (A15201) were purchased from Invitrogen (Thermo Fisher Scientific, Cambridge, Massachusetts, USA); all were stored according to the manufacturer’s instructions.

Du F., Sun L., Chu Y., Li T., Lei C., Wang X., Jiang M., Min Y., Lu Y., Zhao X., Nie Y, & Fan D. (2018). DDIT4 promotes gastric cancer proliferation and tumorigenesis through the p53 and MAPK pathways. Cancer Communications, 38, 45.

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Ang II-induced Cellular Responses

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Ang II was purchased from Sigma-Aldrich (Merck Millipore Corporation, Darmstadt, Germany). PD98059, ERK1/2 inhibitor, and SB203580, p38 MAP kinase inhibitor, were purchased from Invitrogen (Carlsbad, CA, USA). Fimasartan, ARB, was provided by Boryung Pharmaceutical Co., Ltd (Seoul, Korea). Primary antibodies against p53, p16 and β-actin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), and Phospho-ERK1/2, ERK1/2, Phospho-p38 MAPK and p38 MAPK antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). CYR61 antibody was purchased from Abcam. Secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Kim I., Park C.S, & Lee H.Y. (2019). Angiotensin II Type 1 Receptor Blocker, Fimasartan, Reduces Vascular Smooth Muscle Cell Senescence by Inhibiting the CYR61 Signaling Pathway. Korean Circulation Journal, 49(7), 615-626.

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Neural Stem Cell Differentiation

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Dulbecco’s modified Eagle’s medium/F12 (1∶1, DMEM/F12,) and B27 were from Gibco (Grand Island, NY, USA). Epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), trypsin, and dimethyl sulfoxide (DMSO) were purchased from Sigma (St Louis, MO, USA). IGF-1, PD98059, and LY294002 were from Invitrogen (Carlsbad, CA, USA). Both PD98059 and LY294002 were dissolved in DMSO and stored at 50 mM. Rapamycin was from Beyotime (Nantong, CHN). AG1024 was form Selleck (Housten, TX, US). Other reagents are described below.

Zhang X., Zhang L., Cheng X., Guo Y., Sun X., Chen G., Li H., Li P., Lu X., Tian M., Qin J., Zhou H, & Jin G. (2014). IGF-1 Promotes Brn-4 Expression and Neuronal Differentiation of Neural Stem Cells via the PI3K/Akt Pathway. PLoS ONE, 9(12), e113801.

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Hyaluronic Acid and Anti-Cancer Agents

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Hyaluronic acid (HA) was obtained from MP Biomedicals (ThermoFisher Scientific; Fair Lawn, NJ, USA, CAS 9004-61-9) with a molecular weight (MW) of 776.651 g/mol. One chemotherapeutic agent used in other HA-combination therapies studies of head and neck cancers [17 (link)], paclitaxel (PTX) or Taxol, was obtained from ACROS Organics (ThermoFisher Scientific, CAS 33069-62-4) with an MW of 853.9 g/mol. A positive control, the cellular mitogen-activated protein kinase (MAP) kinase (MEK) inhibitor PD98059, was obtained from Invitrogen (ThermoFisher Scientific, Catalog number PHZ1164) with an MW of 267.3 Daltons (Da).

Ringer J., Morrison B, & Kingsley K. (2020). Evaluation of Hyaluronic Acid to Modulate Oral Squamous Cell Carcinoma Growth In Vitro. Journal of Functional Biomaterials, 11(4), 72.

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Elucidating PEMF-Mediated Chondrogenesis Pathways

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After bioinformatics analysis and literature review, ERK and p38-MAPK pathways were located as possible pathways mediating the effects of PEMF on hBMSCs chondrogenesis. To further validate the involvement of ERK and p38-MAPK pathways on chondrogenic differentiation of hBMSCs during PEMF treatments, on day 0, cell pellets were cultured in CIM and received daily PEMF treatment (3 h s/day) for 10 days. Cell pellets from control group were kept at same condition and received no PEMF treatment. Samples were collected on day 1, 3, 7 and 10 and subject to Western blot to analyze the expression and phosphorylation of ERK1/2 and p38 protein.
Pharmacological inhibition study was deployed to investigate the correlation between regulation of ERK and p-38 by PEMF treatment and chondrogenesis of hBMSCs. On day 0, Specific inhibitors of MEK ⁄ ERK (PD98059, 10 μmol/L, Invitrogen™, ThermoFisher Scientific, USA)), p38 (SB20350, 3 μmol/L, Sigma-Aldrich, USA), or a combination of both inhibitors were added into CIM of pellet culture, and cell pellets received daily PEMF treatment (3 h s/day) for 14 days. Samples were collected on day 1 to exam ERK1/2 and p38 activities after pharmacological inhibition through Western blot, and collected on day 7 and 14 for qPCR analysis of chondrogenic marker (Sox9) and hypertrophic marker (RUNX2).

Li Y., Li L., Li Y., Feng L., Wang B., Wang M., Wang H., Zhu M., Yang Y., Waldorff E.I., Zhang N., Viohl I., Lin S., Bian L., Lee W.Y, & Li G. (2022). Enhancing cartilage repair with optimized supramolecular hydrogel-based scaffold and pulsed electromagnetic field. Bioactive Materials, 22, 312-324.

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Piperine-Induced Apoptosis Signaling Pathways

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Piperine (>95% purity), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), dimethylsulfoxide (DMSO), 4,6-diamidino-2-phenylindole (DAPI), rhodamine 123 (Rho-123), and protease inhibitors were purchased from Sigma Chemical (St. Louis, MO). Dulbecco modified Eagle medium (DMEM), fetal bovine serum (FBS), penicillin, Akt inhibitor LY294002, p38 MAPK inhibitor SB203580, JNK1/2 inhibitor SP600125, and ERK1/2 inhibitor PD98059 were from Invitrogen (Grand Island, NY, U.S.A.). Annexin V-FITC Apoptosis Detection kit was from Invitrogen (Carlsbad, CA, U.S.A). BCA Protein Assay Kit was from Pierce (Rockford, IL, U.S.A). Primary antibodies for β-actin, cytosolic cytochrome c, phosphorylated p38 MAPK (p-p38 MAPK) and p38 MAPK, phosphorylated JNK (p-JNK), and cleaved PARP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A). Caspase-3, caspase-8, and caspase-9 colorimetric assay kits are from Kengen Inc, (Beijing, China). All other chemicals and reagents were of analytical grade.

Si L., Yang R., Lin R, & Yang S. (2018). Piperine functions as a tumor suppressor for human ovarian tumor growth via activation of JNK/p38 MAPK-mediated intrinsic apoptotic pathway. Bioscience Reports, 38(3), BSR20180503.

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Intracellular Signaling in L-HLA-Treated DPSCs

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To determine how the intracellular signaling cascade was affected by L-HLA in DPSCs, alternation in specific intracellular kinases in L-HLA-treated DPSCs were assessed. For the ERK, JNK, and p38 signal pathways, 50 µmol/L of PD98059 (Gibco/Invitrogen, Carlsbad, CA, USA), 30 µmol/L of SP600125 (Enzo Life Sciences, Farmingdale, NY, USA), and 10 µmol/L of SB203580 (Gibco/Invitrogen, Carlsbad, CA, USA) were added, respectively, to the culture medium. The cell viabilities were determined for cells treated with or without L-HLA. In addition, to ensure the binding of the prepared L-HLA to HLA receptor on DPSC membranes, MTT assay was performed after adding 0.4 μg/mL anti-CD44 antibody (Thermo Fisher Scientific, Waltham, MA, USA) to the cultured cell system, with and without L-HLA exposure.

Lin C.Y., Kuo P.J., Lin Y.H., Lin C.Y., Lin J.C., Chiu H.C., Hung T.F., Lin H.Y, & Huang H.M. (2022). Fabrication of Low-Molecular-Weight Hyaluronic Acid–Carboxymethyl Cellulose Hybrid to Promote Bone Growth in Guided Bone Regeneration Surgery: An Animal Study. Polymers, 14(15), 3211.

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Apigenin and Methylcellulose Modulate Inflammatory Pathways

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Apigenin (95% purity) and methylcellulose (MC) were obtained from Sigma-Aldrich (St. Louis, Mo), Cerulein (CR) peptide was purchased from Bachem (Torrance, Calif), and PTHrP (1–36) from PolyPeptide Laboratories (San Diego, Calif). Transforming growth factor beta (TGF-β) and tumor necrosis factor alpha (TNF-α) were obtained from R&D Systems (Minneapolis, Minn). PD98059 and LY294002 were purchased from Life Technologies (Carlsbad, Calif) and Selleck Chemicals (Houston, Texas), respectively.

Mrazek A.A., Bhatia V., Falzon M., Spratt H., Chao C, & Hellmich M.R. (2019). Apigenin Decreases Acinar Cell Damage in Pancreatitis. Pancreas, 48(5), 711-718.

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