KRAS-mutant NSCLC cell lines were authenticated and maintained in the recommended medium. Palbociclib (PD0332991) and PD0325901 were supplied by Pfizer. Trametinib, binimetinib, ulixertinib, everolimus, MK2206, erlotinib, LY2874455 and LMN-211904 were obtained from Selleckchem, and bFGF neutralizing antibody was obtained from Sigma. For depletion experiments, pLKO.1 shRNA constructs targeting CDK1, CDK2, CDK4, CDK6, cyclin D1, cyclin D3 and cyclin E (Harvard PlasmID Database) were expressed by lentiviral infection. Palbociclib-resistant cells H358-PR100, H358-PR250, H460-PR500 and H441-PR500 were isolated after the serial addition of increasing concentrations of Palbociclib to culture media. Resistance was confirmed by assessment of proliferation and cell cycle analysis in the presence of Palbociclib. Cell viability assays were performed using CellTiter-Glo (Promega) starting with 1 × 103 cells in 96-well plates.
Pd0325901
Manufactured by Pfizer
Sourced in United States
PD0325901 is a chemical compound that functions as a selective inhibitor of the mitogen-activated protein kinase kinase (MEK) enzyme. It is commonly used as a research tool in cell biology and biochemistry laboratories to investigate the MEK/ERK signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Palbociclib, by Pfizer (2 mentions)
Mek inhibitor, by Pfizer (2 mentions)
Ng nitro l arginine methyl ester, by Merck Group (2 mentions)
Everolimus, by Selleck Chemicals (1 mentions)
Binimetinib, by Selleck Chemicals (1 mentions)
Ulixertinib, by Selleck Chemicals (1 mentions)
Erlotinib, by Selleck Chemicals (1 mentions)
Trametinib, by Selleck Chemicals (1 mentions)
Ly2874455, by Selleck Chemicals (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Celltiter glo, by Promega (1 mentions)
Gdc 0941, by Genentech (1 mentions)
Plakoglobin, by BD (1 mentions)
Iressa, by AstraZeneca (1 mentions)
Fibronectin, by Merck Group (1 mentions)
Fgfr1, by Santa Cruz Biotechnology (1 mentions)
Cytokeratin 18, by Merck Group (1 mentions)
Pd173074, by Pfizer (1 mentions)
Zd1839, by AstraZeneca (1 mentions)
Mk2206, by Bio-Techne (1 mentions)
15 protocols using pd0325901
1
KRAS-mutant NSCLC Cell Line Profiling
2
PD0325901 Formulation and Dosing in Minipigs
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PD0325901 (Pfizer) was obtained from SelleckChem (Houston, TX) and underwent testing at the University of Minnesota’s Institute for Drug Discovery for confirmation of molecular identity and analysis of purity using an acetonitrile/water/formic acid liquid chromatography (LC) conditions. PD0325901 was determined to be >99% pure by LC/mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) analysis. PD0325901 was formulated for oral administration in aqueous 0.5% (w v−1) methylcellulose solution with 0.2% (v/v) polysorbate 80 (Tween 80) to a concentration of 4 mg mL−1 and sonicated to form a suspension. A single oral dose was administered to four wild-type and four NF1 minipigs at 0.79 mg kg−1. The required volume of drug for each animal was based on individual body weight determined on the day of administration.
3
Gamma Secretase Inhibitor Dosing Protocol
4
MTT Assay for Drug Dosage Curves
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The MTT assay was employed to create drug dosage curves. Assays were seeded at a predetermined density. The MEK inhibitor PD0325901 (Pfizer) was dissolved in DMSO to 10 mM and stored at −20°C. Serial dilutions of the drug in media were prepared and 100 µl/well added. The assay was then performed in the same manner as above with varying incubation periods (24, 48, 72 or 96 hours) prior to addition of the labeling reagent.).
5
Acquisition and Preparation of PF-384 and PD-901
6
In Vitro and In Vivo Inhibition of Signaling Pathways
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Two signal transduction pathway inhibitors were used in this study. PD0325901 (Pfizer; a kind gift from Martin McMahon, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA) is a selective and non ATP-competitive MEKi, and GDC-0941 (a kind gift from Genentech) is a highly selective class I PI3Ki specific for p110α and p110β subunits (Folkes et al., 2008 (link)). For in vitro primary myoblast culture inhibitor experiments, 1 µM final concentration of MEKi PD0325901 or PI3Ki GDC-0941 was added to low-serum DM until cells were harvested. For in vivo inhibitor treatment, 3-month-old HrasG12V and 3-month-old WT littermates were administered either MEK inhibitor at 12.5 mg/kg (Trejo et al., 2012 (link)) or PI3K inhibitor at 75 mg/kg once a day (Folkes et al., 2008 (link)) for 28 days by oral gavage. Control mice were given vehicle only. Each study arm had five mice per group, except for the PI3Ki arm, which had three mice. The inhibitor compounds were formulated in 0.5% (w/v) hydroxypropyl methyl cellulose, which also served as the vehicle (Sigma-Aldrich).
7
PF-502 and PD-901 Preparation Protocols
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PF-04691502 (PF-502) and PD-0325901 (PD-901) were provided by Pfizer Inc. (San Diego, CA). For in vitro work, both agents were dissolved in 100% DMSO at a concentration of 10 mM. For in vivo studies, PF-502 was suspended in 0.5% methylcellulose in water, and PD-901 was suspended in 0.5% hydroxypropyl methylcellulose, 0.2% tween-80 in water.
8
Epithelial-Mesenchymal Transition Signaling
9
Gamma Secretase Inhibitor Dosing Protocol
10
Gamma Secretase Inhibitor Dosing Protocol
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Gamma secretase inhibitor (GSI,MRK-003; Merck) was made fresh weekly and dosed at 300 mg/kg in 0.5% methocel by oral gavage (Lewis et al., 2007 (link); Sparey et al., 2005 (link)). For pathology, we dosed mice once weekly for 4 weeks and sacrificed 6 hr after the last dose (n = 5 doses). For flow analysis, we dosed mice once weekly for 1 week and sacrificed mice 6 hr after the last dose (n = 2 doses). Fresh solution of L-NAME (L-NG-nitroarginine methyl ester, 100 μM in 1× PBS; Sigma-Aldrich) was administered daily at 0.4 mg/kg. For pathology and flow analysis, mice were injected intraperitoneally (i.p.) daily for 7 days and sacrificed 6 hr after the last dose. MEK inhibitor (PD0325901; Pfizer) was made fresh weekly and dosed at 1.5 mg/kg/day in 0.5% methocel/0.2% Tween 80, by oral gavage. For pathology, we dosed mice every day for 3 weeks. For flow analysis, we dosed mice every day for 7 days and sacrificed 6 hr after the last dose.
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