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Pimasertib

Manufactured by Selleck Chemicals
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Pimasertib is a small molecule inhibitor that targets the MEK1 and MEK2 proteins, which are part of the MAPK/ERK signaling pathway. It is used in research applications to study the role of the MAPK/ERK pathway in various cellular processes and disease models.

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Lab products found in correlation

Trametinib, by Selleck Chemicals (9 mentions) Selumetinib, by Selleck Chemicals (5 mentions) Binimetinib, by Selleck Chemicals (3 mentions) Refametinib, by Selleck Chemicals (3 mentions) Azd8330, by Selleck Chemicals (3 mentions) Tak 733, by Selleck Chemicals (3 mentions) Etoposide, by Thermo Fisher Scientific (3 mentions) Bortezomib, by Thermo Fisher Scientific (3 mentions) Q vd oph, by Thermo Fisher Scientific (3 mentions) Dabrafenib, by Selleck Chemicals (3 mentions) Lds sample buffer, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Cobimetinib, by Selleck Chemicals (2 mentions) Ro5126766, by Selleck Chemicals (2 mentions) Pd0325901, by Axon Medchem (2 mentions) Vinblastine, by Selleck Chemicals (2 mentions) Camptothecin, by Selleck Chemicals (2 mentions) Oligomycin, by Merck Group (2 mentions) A 1155463, by Selleck Chemicals (2 mentions)

17 protocols using pimasertib

1

Isolation and Culture of Mouse Cortical Neurons

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Mouse cortical neurons were prepared from E16.5 FVB/N mice and plated in poly-D-lysine coated 12-well plates (5×105 per well) in Neurobasal medium supplemented with GlutaMAX (Invitrogen), B-27, antibiotics (Penicillin/Streptomycin). Half of the media was changed with fresh media every 3–4 days. For drug treatment, DMSO (Sigma), pimasertib (2 μM, Selleckchem), or selumetinib (0.4 μM, Selleckchem) was applied to the media starting at days in vitro (DIV) 1. For shRNA virus infection experiments, neurons were infected with lentiviruses at DIV 1 with the multiplicity of infection at 10. Neurons were harvested at DIV 15–18 and lysed with RIPA buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 1 % NP-40, 0.5% sodium deoxycholate, 0.1% SDS) supplemented with phosphatase and protease inhibitors (GenDEPOT). Lysates were cleared by centrifugation (20 min, 15,000 rpm, 4°C) followed by protein quantification via BCA assay (Pierce). LDS Sample buffer (Invitrogen) with a reducing agent was added to each lysate followed by a 10 min incubation at 95°C. Samples were spun down and electrophoresed on a 4–12% Bis-Tris gel, transferred to a nitrocellulose membrane and blocked for one hour with 5% non-fat milk prior to primary antibody incubation. Unless otherwise indicated, the samples used for western blot were from whole lysates.
Wang L., Adamski C.J., Bondar V.V., Craigen E., Collette J.R., Pang K., Han K., Liu Z., Sifers R.N., Holder JL J.r, & Zoghbi H.Y. (2019). A kinome-wide RNAi screen identifies ERK2 as a druggable regulator of Shank3 stability. Molecular psychiatry, 25(10), 2504-2516.
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2

MEKi Culturing of Naive hESCs

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The following MEKi were used to culture naïve hESCs at the concentrations indicated in the manuscript: PD0325901 (Axon Medchem), Selumetinib (Selleck Chemicals), Binimetinib (Selleck Chemicals), Trametinib (Selleck Chemicals), Pimasertib (Selleck Chemicals), Refametinib (Selleck Chemicals), TAK-733 (Selleck Chemicals), RO5126766 (Selleck Chemicals), Cobimetinib (Selleck Chemicals), AZD8330 (Selleck Chemicals).
Stefano B.D., Ueda M., Sabri S., Brumbaugh J., Huebner A.J., Sahakyan A., Clement K., Clowers K.J., Erickson A.R., Shioda K., Gygi S.P., Gu H., Shioda T., Meissner A., Takashima Y., Plath K, & Hochedlinger K. (2018). Reduced MEK inhibition preserves genomic stability in naïve human ES cells. Nature methods, 15(9), 732-740.
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3

Evaluation of Bioactive Compounds in Cell Screening

Cited 1 time
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Camptothecin (Cat# S1288), vinblastine (Cat# S1248), pimasertib (Cat# S1457), trametinib (Cat# S2673), ABT-737 (Cat# S1002), A-1155463 (Cat# S7800), and nutlin-3 (Cat# S1061) were purchased from Selleck Chemicals (Houston, TX). Bortezomib (Cat# NC0587961), etoposide (Cat# ICN19391825), MG-132 (Cat# 17-485), and Q-VD-OPh (Cat# OPH00101M) were purchased from Thermo Fisher Scientific. N-acetylcysteine (Cat# A8199), thapsigargin (Cat# T9033), tunicamycin (Cat# T7765), paclitaxel (Cat# T7191), JNK Inhibitor VIII (Cat# 420135), 2-deoxyglucose (Cat# D8375), oligomycin (Cat# O4876), and cycloheximide (Cat# C7698) were obtained from Sigma-Aldrich (St. Louis, MO). S63845 (Cat#21131) was obtained from Cayman Chemical (Ann Arbor, MI). Staurosporine (Cat# A8192) was obtained from ApexBio (Houston, TX). Erastin was the kind gift of Brent Stockwell (Columbia University). Erastin2 (compound 35MEW28 in Dixon et al., [2014 (link)]) and ML162 (CAS: 1035072-16-2) were synthesized by Acme Bioscience (Palo Alto, CA). Chemical screening was conducted as described below; the library of 261 bioactive compounds was obtained from Selleck Chemicals (Cat# L2000).
Inde Z., Forcina G.C., Denton K, & Dixon S.J. (2020). Kinetic Heterogeneity of Cancer Cell Fractional Killing. Cell reports, 32(1), 107845.
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4

Targeting BRAF, NRAS, and cKIT Mutants

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The BRAF inhibitor Dabrafenib, used for BRAF mutated cells, and the MEK inhibitor Pimasertib, used for NRAS mutated cells, were from Selleck Chemicals (Houston, TX, USA). The tyrosine kinase inhibitor Dasatinib, used for cKIT mutated cells, was from Bristol-Myers (New York, NY, USA). Bufalin was from PhytoLab (Vestenbergsgreuth, Germany). Methyl-β-cyclodextrin is from Merck (Burlington, MA, USA).
Soumoy L., Genbauffe A., Mouchart L., Sperone A., Trelcat A., Mukeba-Harchies L., Wells M., Blankert B., Najem A., Ghanem G., Saussez S, & Journe F. (2024). ATP1A1 is a promising new target for melanoma treatment and can be inhibited by its physiological ligand bufalin to restore targeted therapy efficacy. Cancer Cell International, 24, 8.
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5

Modulating Intracellular Signaling Pathways in Salmonella Infection

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Cells were infected with S. Tm at MOI of 10 for 1 h. Cells were then rinsed with PBS and replaced with DMEM containing 50 μg/ml gentamycin and one of the inhibitors or activators which are listed as follows: Cdc42 inhibitor, 5 µM ML141 (MedChem Express, HY-12755, CN); MEK1/2 inhibitors, 1 µM Trametinib (S2673), 1 µM RDEA119 (S1089), 1 µM Pimasertib (S1475), 1 µM AZD8330 (S2134), 1 µM TAK-733 (S2617), 1 µM BI-847325 (S7843), 10 µM U0126 (S1102) from Selleck chemicals (US); Cdc42 activator, 100 ng/ml Bradykinin (TargetMol, TP1277, CN); MEK activator, 100 ng/ml human EGF (Novoprotein, CA). Cells continued to incubate for an additional 24 h.
Zhao S., Xu Q., Cui Y., Yao S., Jin S., Zhang Q., Wen Z., Ruan H., Liang X., Chao Y., Gong S., Sansonetti P., Wei K., Tang H, & Jiu Y. (2023). Salmonella effector SopB reorganizes cytoskeletal vimentin to maintain replication vacuoles for efficient infection. Nature Communications, 14, 478.
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6

High-throughput Screening of Ferroptosis Compounds

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ML162 was synthesized by Acme (Palo Alto, CA). Erastin2 (Cat# 27087) and deferoxamine (Cat# 14595) were obtained from Cayman Chemicals (Ann Arbor, MI). Palbociclib (Cat# S1116), pimasertib (Cat# S1475), and 1S,3R-RSL3 (simply, RSL3) (Cat# S8155) were obtained from Selleck Chemicals (Houston, TX). Doxycycline (Dox) (Cat# D3447), ML210 (Cat# SML0521), ferrostatin-1 (Cat# SML0583) and propidium iodide (Cat# P4170) were obtained from Sigma-Aldrich (St. Louis, MO). Abemaciclib (Cat# HY-16297A) was obtained from MedChem Express (Princeton, NJ). Bortezomib (Cat# NC0587961), Q-VD-OPh (Cat# OPH00101M), etoposide (Cat# 12-261-00), DAPI (Cat# D1306) and C11 BODIPY 581/591 were from Thermo Fisher Scientific (Waltham, MA). Compound 28 was synthesized as described47 and provided as a gift by P. Beltran (Ferro Inc). Dox was dissolved in deionized water and stored at −20°C. C11 BODIPY 581/591 was dissolved into methanol and stored at −20°C. All other compounds were dissolved in dimethyl sulfoxide (DMSO) and then stored at −20°C. The compound library (Cat# L1700) was obtained from Selleck Chemicals8 (link).
Rodencal J., Kim N., Li V.L., He A., Lange M., He J., Tarangelo A., Schafer Z.T., Olzmann J.A., Sage J., Long J.Z, & Dixon S.J. (2023). A Cell Cycle-Dependent Ferroptosis Sensitivity Switch Governed by EMP2. bioRxiv.
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7

Evaluating TEAD-dependent BRAF/MEK inhibition

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Overall, 200,000 cells were plated per six wells. Cells were transfected to KD all TEADs as described above. On day 3, 10,000 or 3,000 cells were seeded in 96 wells in triplicates. On day 4, variable concentrations ranging between 60 and 0.05 μM of BRAF inhibitor (PLX4032, Selleck Chemicals), MEK inhibitor (Pimasertib, Selleck Chemicals) were added. Alternatively, BRAF or MEK inhibitors were added at concentrations approximating the predicted IC50 value for each individual culture to measure potential additive effect of the TEAD KD. Cells treated with DMSO were used as a control and they signify maximum cell viability. Forty-eight or seventy-two hours after stimulation, cell viability was measured as described above. All measurements were carried out in triplicates. Final data points are averages of at least three biological replicates. IC50 shift curves were generated using the appropriate analysis protocol from Graphpad Prism.
Verfaillie A., Imrichova H., Atak Z.K., Dewaele M., Rambow F., Hulselmans G., Christiaens V., Svetlichnyy D., Luciani F., Van den Mooter L., Claerhout S., Fiers M., Journe F., Ghanem G.E., Herrmann C., Halder G., Marine J.C, & Aerts S. (2015). Decoding the regulatory landscape of melanoma reveals TEADS as regulators of the invasive cell state. Nature Communications, 6, 6683.
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8

Targeting BRAF, NRAS, and cKIT Mutations

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The BRAF inhibitor Dabrafenib, used for BRAF mutated cells, and the MEK inhibitor Pimasertib, used for NRAS mutated cells, were from Selleck Chemicals (Houston, TX, USA). The tyrosine kinase inhibitor Dasatinib, used for cKIT mutated cells, was from Bristol-Myers (New York, NY, USA).
Soumoy L., Schepkens C., Krayem M., Najem A., Tagliatti V., Ghanem G.E., Saussez S., Colet J.M, & Journe F. (2020). Metabolic Reprogramming in Metastatic Melanoma with Acquired Resistance to Targeted Therapies: Integrative Metabolomic and Proteomic Analysis. Cancers, 12(5), 1323.
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9

Zebrafish Tailfin Area Modulation

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Zebrafish embryos were collected at 1 dpf and placed in a 40-μm cell strainer (Thermo Fisher 08–771-2) in a 6-well dish (Fisher 08–772-1B) in 6 ml E3 water. Twenty embryos were used per well. Zebrafish embryos were treated with the indicated compounds: insulin and IGF1 receptor antagonists BMS-754807 (Sigma-Aldrich BM0003–5MG) at 7.5 μM and NVP-AEW541(Selleckchem S1034) at 60 μM, PI3K inhibitor LY294002 (Sigma-Aldrich L9908–1MG) at 15 μM, RAF/MEK inhibitor CH5126766 (Selleckchem S7170) at 1 μM, MEK inhibitor Pimasertib (Selleckchem S1475) at 1 μM, MEK inhibitor Refametinib (Selleckchem #S1089) at 1 μM, MEK inhibitor Trametinib (Selleckchem S2673) at 200 nM, and SOS1 inhibitor Bl-3406 at 1 μM (MedChemExpress HY-125817). Compound stocks were kept at 1,000× in DMSO. Treatment started at 1 dpf and was reapplied at 2 dpf. Zebrafish were imaged for tailfin area at 3 dpf.
Weiss J.M., Hunter M.V., Cruz N.M., Baggiolini A., Tagore M., Ma Y., Misale S., Marasco M., Simon-Vermot T., Campbell N.R., Newell F., Wilmott J.S., Johansson P.A., Thompson J.F., Long G.V., Pearson J.V., Mann G.J., Scolyer R.A., Waddell N., Montal E.D., Huang T.H., Jonsson P., Donoghue M.T., Harris C.C., Taylor B.S., Xu T., Chaligné R., Shliaha P.V., Hendrickson R., Jungbluth A.A., Lezcano C., Koche R., Studer L., Ariyan C.E., Solit D.B., Wolchok J.D., Merghoub T., Rosen N., Hayward N.K, & White R.M. (2022). Anatomic position determines oncogenic specificity in melanoma. Nature, 604(7905), 354-361.
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10

Evaluating BRAF and MEK Inhibitor Efficacy

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Cells were plated at 10,000 or 3,000 cells per well in a 96-well plate depending on the length of the experiment. After 24 h, a dilution series for BRAF (PLX4032, Selleck Chemicals) or MEK inhibitor (Pimasertib, Selleck Chemicals) was created ranging between 20 and 0.05 μM. Forty-eight or seventy-eight hours after stimulation, cell viability was measured. Measurements were normalized using cells treated with dimethylsulphoxide (DMSO) control as maximal viability. Each condition was performed in triplicates and final data points are the average of at least duplicate biological replicates. Curves were fitted and IC50 calculated using a nonlinear regression analysis in Graphpad Prism (version 6.0 for mac OS X, GraphPad Software, San Diego, CA, USA, www.graphpad.com).
Verfaillie A., Imrichova H., Atak Z.K., Dewaele M., Rambow F., Hulselmans G., Christiaens V., Svetlichnyy D., Luciani F., Van den Mooter L., Claerhout S., Fiers M., Journe F., Ghanem G.E., Herrmann C., Halder G., Marine J.C, & Aerts S. (2015). Decoding the regulatory landscape of melanoma reveals TEADS as regulators of the invasive cell state. Nature Communications, 6, 6683.
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