Pd98059

Manufactured by Cell Signaling Technology

Sourced in United States, Germany, United Kingdom, Japan, China

PD98059 is a potent and selective inhibitor of the mitogen-activated protein kinase kinase (MEK) enzymes, MEK1 and MEK2. It blocks the activation of the extracellular signal-regulated kinase (ERK) pathway, which is involved in cell proliferation and differentiation.

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Close spelling variants of this product

PD98059 (PD) (6 citations) MEK1/2 inhibitor PD98059 (3 citations) PD98059 (9900S), (2 citations)

226 protocols using pd98059

Investigating Menthol and Formaldehyde Effects on Neurons

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After allowing the neurites to freely extend for 48 hours, the DRG neurons were pretreated with capsazepine (a TRPV1 antagonist, 10 μM) (Sigma-Aldrich) for 30 minutes. Thereafter, the neurons were treated and cultured for 24 hours as follows: (1) formaldehyde group (n = 5), DRG neurons were exposed to formaldehyde (Sigma-Aldrich) 10 μM; (2) menthol group (n = 5), DRG neurons were exposed to menthol (Sigma-Aldrich) 300 μM; (3) PD98059 + formaldehyde group (n = 5), PD98059 (an ERK1/2 inhibitor, 10 μM) (Cell Signaling Technology, Danvers, MA, USA) was added 30 minutes before formaldehyde (10 μM) exposure; (4) PD98059 + menthol group (n = 5), PD98059 (10 μM) was added 30 minutes prior to menthol (300 μM) exposure; (5) control group (n = 5), DRG neurons treated only with the TRPV1 receptor antagonist capsazepine (10 μM). Because TRPA1 and TRPV1 channels interact in neurons (Ruparel et al., 2011), capsazepine, a TRPV1 antagonist, was used to block the effects of TRPV1.

Wang X.L., Cui L.W., Liu Z., Gao Y.M., Wang S., Li H., Liu H.X, & Yu L.J. (2019). Effects of TRPA1 activation and inhibition on TRPA1 and CGRP expression in dorsal root ganglion neurons. Neural Regeneration Research, 14(1), 140-148.

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Cell Line Cultivation and SFN Treatment

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SK-1 cells and A549 cells from Cell Resource Center, Peking Union Medical College (CRC/PUMC) were cultured in DMEM/F-12 medium (Lifetechnologies, Shanghai, China) supplemented with 10% fetal bovine serum(Invitrogen), and 100 U/ml penicillin/streptomycin (Solarbio, Beijing, China) at 37℃in 5% CO₂, maintaining cells logarithmic growth. Cells were washed with PBS three times and incubated in DMEM/F-12 medium supplemented with 10% FBS and 100U/ml penicillin/streptomycin and SFN (Sigma) for 24h. MG132 (Sigma) and PD98059 (Cell Signaling Technologies) had been added before SFN was added into F-12 medium according to PD98059 product specification.

Geng Y., Zhou Y., Wu S., Hu Y., Lin K., Wang Y., Zheng Z, & Wu W. (2017). Sulforaphane Induced Apoptosis via Promotion of Mitochondrial Fusion and ERK1/2-Mediated 26S Proteasome Degradation of Novel Pro-survival Bim and Upregulation of Bax in Human Non-Small Cell Lung Cancer Cells. Journal of Cancer, 8(13), 2456-2470.

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Osteogenic Differentiation of hMSCs

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3 × 10⁵ hMSCs were seeded onto 8 mm discs of CG-GAG and MC-GAG scaffolds in proliferation media. 24 h after seeding, media was switched to osteogenic differentiation media consisting of 10 mM β-glycerophosphate, 50 μg/mL ascorbic acid, and 0.1 μM dexamethasone. For inhibitor studies, scaffolds were treated or untreated with dorsomorphin homologue 1 (DMH1; Sigma-Aldrich) or PD98059 (Cell Signaling Technologies, Beverly, MA) separately, all at a concentration of 50 μM. Fresh DMH1 and PD98059 were added to each media change every 3 days.

Ren X., Zhou Q., Foulad D., Dewey M.J., Bischoff D., Miller T.A., Yamaguchi D.T., Harley B.A, & Lee J.C. (2019). Nanoparticulate Mineralized Collagen Glycosaminoglycan Materials Directly and Indirectly Inhibit Osteoclastogenesis and Osteoclast Activation. Journal of tissue engineering and regenerative medicine, 13(5), 823-834.

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Regulation of NSCs by hUC-MSCs-CM

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Neurosphere cultures were prepared from newborn rat cortex and grown in serum-free medium containing EGF and bFGF (both 20 ng/ml), For the differentiation assay, neurospheres were dissociated and plated on poly-lysine coated coverslips in the absence or presence of hUC-MSCs-CM for 3 d before analyses.^{47 (link)}To verify whether the ERK-CREB pathway is involved in the regulation of hUC-MSCs-CM (without any other additional factors) on NSCs, NSCs was pretreated with DMEM/F12 (Gibco) for 12 h. Next, NSCs was pretreated with 20 μM of PD98059 (#9900, Cell Signaling Technology) for 1 h in PD98059 group. Then, the three groups were severally treated with DMEM/F12, hUC-MSCs-CM and 20 μM of PD98059 in hUC-MSCs-CM for 6 h to evaluate the phosphorylation of ERK and CREB, and 3 days to proliferation observation.

Cao N., Liao T., Liu J., Fan Z., Zeng Q., Zhou J., Pei H., Xi J., He L., Chen L., Nan X., Jia Y., Yue W, & Pei X. (2017). Clinical-grade human umbilical cord-derived mesenchymal stem cells reverse cognitive aging via improving synaptic plasticity and endogenous neurogenesis. Cell Death & Disease, 8(8), e2996-.

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Investigating NRG-1β Signaling Pathways

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The established cocultures were randomly divided into the following groups with the corresponding treatment: (1) NRG-1β: The coculture was treated with NRG-1β (20 nmol/L, Peprotech, Rocky Hill, NJ). (2) NRG-1β + PD98059: ERK1/2 inhibitor PD98059 (10 μmol/L, Cell Signaling Technology) was applied 30 min prior to NRG-1β (20 nmol/L) treatment. (3) NRG-1β + LY294002: PI3K inhibitor LY294002 (10 μmol/L, Invitrogen) was applied 30 min prior to NRG-1β (20 nmol/L) treatment. (4) NRG-1β + AG490: JAK2 inhibitor AG490 (10 μmol/L, Invitrogen) was applied 30 min prior to NRG-1β (20 nmol/L) treatment. (5) Control group: The cells were cultured continuously in media. All aforementioned incubation conditions were maintained in 37 °C and 5% CO₂ environment with the corresponding agents for 4 days with media change every 2 days.

Qiao Y., Cong M., Li J., Li H, & Li Z. (2018). The effects of neuregulin-1β on intrafusal muscle fiber formation in neuromuscular coculture of dorsal root ganglion explants and skeletal muscle cells. Skeletal Muscle, 8, 29.

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Modulating Autophagy and MAPK in Embryos

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st24 embryos were dechorionated and incubated for 24 h in 50 nM Bafilomycin A (Sigma), 25 μM PD98059 (Cell Signaling), or 6 μM HA14‐1 (Sigma). Drugs were diluted in 3% DMSO/embryo medium. Medium was refreshed twice a day. Control embryos were grown in 3% DMSO/embryo medium.

Indrieri A., Carrella S., Romano A., Spaziano A., Marrocco E., Fernandez‐Vizarra E., Barbato S., Pizzo M., Ezhova Y., Golia F.M., Ciampi L., Tammaro R., Henao‐Mejia J., Williams A., Flavell R.A., De Leonibus E., Zeviani M., Surace E.M., Banfi S, & Franco B. (2019). miR‐181a/b downregulation exerts a protective action on mitochondrial disease models. EMBO Molecular Medicine, 11(5), e8734.

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LPA-Induced Signaling Pathway Inhibition

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1-Oleoyl LPA (18 : 1 LPA) was purchased from Avanti Polar Lipids (Birmingham, AL, USA). Inhibitor of phosphoinositide 3-kinase (PI3K), LY290042, and inhibitor of mitogen-activated protein kinase (MAPK), PD98059, were from Cell Signaling (Beverly, MA, USA). Rho kinase inhibitor, Y-27632, was from Biomol (Beverly, MA, USA). Hela cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 100 mL/L fetal bovine serum (FBS), streptomycin (100 mg/L), and penicillin (100 KU/L) at 37°C in 50 mL/L CO₂ incubator. Cells were serum starved for 12 hours before LPA treatment.

Sui Y., Yang Y., Wang J., Li Y., Ma H., Cai H., Liu X., Zhang Y., Wang S., Li Z., Zhang X., Wang J., Liu R., Yan Y., Xue C., Shi X., Tan L, & Ren J. (2015). Lysophosphatidic Acid Inhibits Apoptosis Induced by Cisplatin in Cervical Cancer Cells. BioMed Research International, 2015, 598386.

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Human Bronchial Epithelial Cell Culture

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The human bronchial epithelial cell line 16HBE14o-, an immortalized human bronchial epithelial cell line, was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing L-glutamine, 10% FBS and penicillin (100 U/ml) and streptomycin (100 µg/ml). The tissue culture plates were coated using human fibronectin (1 mg/ml), collagen I bovine (3 mg/ml), and bovine serum albumin (1 mg/ml). All the cells used in the experiments were between passages 25 and 50, and were grown and maintained at 37 °C in a 5% CO₂ humidified incubator. Primary human bronchial epithelial cells (HBECs) were isolated from excess donor tissue obtained at the time of lung transplantation under a protocol approved by UNC Medical School IRB. Primary HBECs were cultured as previously described and studied when fully differentiated [6 (link), 24 (link)]. Lactacystin, UO126, UO124, SB203580, and SP600125 were from Calbiochem (La Jolla, CA). PD98059 was purchased from Cell Signaling Technology. The proteasome inhibitor MG132, and the lysosomal inhibitors, leupeptin and chloroquine, were purchased from Sigma-Aldrich (St. Louis, MO).

Xu X., Balsiger R., Tyrrell J., Boyaka P.N., Tarran R, & Cormet-Boyaka E. (2015). Cigarette smoke exposure reveals a novel role for the MEK/ERK1/2 MAPK pathway in regulation of CFTR. Biochimica et biophysica acta, 1850(6), 1224-1232.

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Hedgehog Signaling Pathway Modulation

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Cells were treated 24 h with SMO agonists 200 and 400 nM SAG (566660, Calbiochem) and 3 µM purmorphamine (4551, Tocris bioscience), or antagonist 5 µM vismodegib (S1082, Selleckchem), or kinase inhibitors: 10 µM GF109203X (ALX-270-049, Enzo Life Sciences), 10 µM SB431542 (1614, Tocris bioscience), 20 µM PD98059 (9900, Cell Signaling Technologies), 5 µM SP600125 (270-339-M005, Alexis Biochemicals), 10 µM SB216763 (1616, Tocris bioscience), 200 nM Rapamycin (R0161, LKT Laboratories), 50 µM LY294002 (70920, Cayman Chemical), 5 µM AG1478 (270-036-M001, Alexis Biochemicals), 3 µM Gö6976 (12060S, Cell Signaling Technologies), 10 µM Rottlerin (1610, Tocris bioscience), 20 µM ZIP (2549, Tocris bioscience, kind gift from Prof. Anthony Oro, Stanford University School of Medicine).
To induce cilia in proliferating condition, hTERT-RPE1 cells were treated for 16 h with 50 nM cytochalasin D (250255, MERCK).

Park H.S., Papanastasi E., Blanchard G., Chiticariu E., Bachmann D., Plomann M., Morice-Picard F., Vabres P., Smahi A., Huber M., Pich C, & Hohl D. (2021). ARP-T1-associated Bazex–Dupré–Christol syndrome is an inherited basal cell cancer with ciliary defects characteristic of ciliopathies. Communications Biology, 4, 544.

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Chemical Reagents for Signaling Pathways

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Chemical reagents U0126 (Cell Signaling Technology, #9903), PD98059 (Cell Signaling Technology, #9900), Y-27632 (Cell Signaling Technology, #13624), SB431542 (Selleckchem, S1067), CCG1423 (Selleckchem, S7719), verteporfin (Sigma, SML0534). VS-6064 (Selleckchem, S7654), PF431396 (Selleckchem, S7644), PF562271 (Selleckchem, S2890), and Vanadate (New England Biolabs, P0758) were used at doses and times indicated in figure legends. All compounds were dissolved in DMSO.

Sato T., Verma S., Andrade C.D., Omeara M., Campbell N., Wang J.S., Cetinbas M., Lang A., Ausk B.J., Brooks D.J., Sadreyev R.I., Kronenberg H.M., Lagares D., Uda Y., Pajevic P.D., Bouxsein M.L., Gross T.S, & Wein M.N. (2020). A FAK/HDAC5 signaling axis controls osteocyte mechanotransduction. Nature Communications, 11, 3282.

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