ET-1 was from Merck Millipore, prepared as a 200 μM stock in 5% acetic acid and applied at 100 nM. Phenylephrine (PE) was from Sigma-Aldrich, prepared as a 10 mM stock in water and applied at 10 μM. Inhibitor or vehicle was applied 30 min prior to cell stimulation with agonist. BQ123 was from Merck Millipore, prepared in water and was used at 1 μM. Prazosin was from Tocris, prepared in DMSO and was applied at 100 nM. Dynole (Dynole 34-2) was from Tocris, was prepared in Ethanol and was applied at 10 μM. PD184352 (PD) was from Sigma-Aldrich, prepared in DMSO and used at 10 μM. Actinomycin D (Act D) was from Tocris, prepared in DMSO and applied at 2 μg/ml. Controls were exposed to vehicle.
Pd184352 pd
Manufactured by Merck Group
Sourced in United States
PD184352 (PD) is a laboratory compound used in research applications. It functions as a potent and selective inhibitor of the mitogen-activated protein kinase (MAPK) pathway. The core function of PD184352 (PD) is to modulate cellular signaling processes involved in various biological systems.
Automatically generated - may contain errors
2 protocols using pd184352 pd
1
Agonist-Induced Signaling Pathway Modulation
2
Regulation of PEDF Expression by PKC, cAMP, MAPK, and PI3K Pathways
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Activators -serum-starved (0 and 0.1% CS-FBS) LH-15 cells and PHGC, respectively, were incubated with i) PKC activator -Phorbol 12-myristate 13-acetate (PMA; P1585, Sigma; dissolved in DMSO), for 1 h; ii) human amphiregulin (AREG; 10558-HNAE, Sino Biological, Inc., Beijing, People's Republic China; dissolved in DDW) that interacts with the epidermal growth factor receptor (EGFR) and promotes cell growth, for 4 (LH-15 cells) or 16 (PHGC) hours. PEDF level (protein and mRNA) was detected at the end of the incubation period. Inhibitors -serum-starved (0.1% CS-FBS) LH-15 cells were incubated for 30 min with i) selective PKC inhibitor GF 109203X (GF, G2911, Sigma); ii) potent cAMP-dependent protein kinase inhibitor H89 dihydrochloride hydrate (H89, B1427, Sigma); iii) MEK (MKK1; MAPK kinase) inhibitor PD184352 (PD, PZ0181 Sigma); iv) phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin (W1628, Sigma) or v) EGFR inhibitor AG18 (AG; S8009, selleckchem.com, Houston, TX, USA). All inhibitors were dissolved in DMSO. Culture media were replaced after stimulation with fresh starvation media (0.1% CS-FBS), supplemented with hCG (1 IU) for additional 6 h of incubation, after which PEDF level (protein and mRNA) was detected.
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