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Skepinone l

Manufactured by Selleck Chemicals
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Skepinone-L is a laboratory reagent that functions as a selective inhibitor of the PI3 kinase enzyme. It is used in research applications to study the role of PI3 kinase signaling pathways.

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Lab products found in correlation

Jnk in 8, by Selleck Chemicals (1 mentions) Go6983, by Selleck Chemicals (1 mentions) Ipatasertib, by Selleck Chemicals (1 mentions) Cyclosporin a, by Selleck Chemicals (1 mentions) R0 31 8220, by Selleck Chemicals (1 mentions) Fk506, by Selleck Chemicals (1 mentions) Mk 2206, by Selleck Chemicals (1 mentions) Celltiter glo 2, by Promega (1 mentions) Plx3397, by Apexbio (1 mentions) Flat clear bottom white polystyrene poly d lysine coated microplates, by Corning (1 mentions) Dabrafenib, by Selleck Chemicals (1 mentions) Cobimetinib, by Selleck Chemicals (1 mentions) Trametinib, by Selleck Chemicals (1 mentions) Vemurafenib, by Selleck Chemicals (1 mentions) Anisomycin, by Fujifilm (1 mentions) Phos tag ligand, by Fujifilm (1 mentions) Bi d1870, by Abcam (1 mentions) β actin c4, by Santa Cruz Biotechnology (1 mentions) Birb 796, by Selleck Chemicals (1 mentions) Gsk2334470, by MedChemExpress (1 mentions)

5 protocols using skepinone l

1

Elucidating Splicing Regulation by TCR Signaling

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To elucidate the roles of signal transduction proteins to regulate splicing changes, each protein known to be regulated downstream of TCR signaling was targeted by at least two different inhibitors. Human primary CD4+ CD45RO- T cells were incubated with individual inhibitors for 1 hr and stimulated with bound anti-CD3 and soluble anti-CD28 antibodies for 48 hr. Cells were then harvested for RNA purification and low-cycle RT-PCR analysis for splicing quantifications. The inhibitors utilized are as follows: PCKi: R0-31-8220 (Selleckchem: S7207) and Go6983 (Selleckchem: S2911), p38i: SB20350 (Selleckchem: S1076) and Skepinone-L (Selleckchem: S7214), NFATi: Cyclosporin A (Selleckchem: S2286) and FK506 (Selleckchem: S5003), AKTi: MK-2206 (Selleckchem:S1078) and Ipatasertib (Selleckchem:S2808), JNKi: SP600215 (Selleckchem: S1460) and JNK-IN-8 (Selleckchem: S4901) and Tanzisertib (Selleckchem: S8490).
Blake D., Radens C.M., Ferretti M.B., Gazzara M.R, & Lynch K.W. (2022). Alternative splicing of apoptosis genes promotes human T cell survival. eLife, 11, e80953.
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2

Microglia Differentiation and Inhibition Assay

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First, 10,000 iTF-iPSCs were seeded into 96-well Flat Clear Bottom White Polystyrene Poly-d-Lysine Coated Microplates (Corning; Cat. No. 3843) and differentiated into iTF-Microglia. For CSF1R inhibition, cells were treated with the CSF1R inhibitor PLX3397 at day 8 (ApexBio; Cat. No. B5854) at indicated concentrations or dimethylsulfoxide control for 24 h before performing the CellTiter-Glo 2.0 (Promega; Cat. No. G9242) assay according to the manufacturer’s instructions. For MAPK14 inhibition, cells were treated with the MAPK14 inhibitor Skepinone-L at day 8 (Selleckchem; Cat. No. 1221485831) at indicated concentrations or dimethylsulfoxide control for 24 h before performing the CellTiter-Glo 2.0 assay. Luminescence signal was recorded with the M5 plate reader (SpectroMax) using SoftMax Pro 6.5.1.
Dräger N.M., Sattler S.M., Huang C.T., Teter O.M., Leng K., Hashemi S.H., Hong J., Aviles G., Clelland C.D., Zhan L., Udeochu J.C., Kodama L., Singleton A.B., Nalls M.A., Ichida J., Ward M.E., Faghri F., Gan L, & Kampmann M. (2022). A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states. Nature Neuroscience, 25(9), 1149-1162.
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3

Pharmacological Inhibition of Signaling Pathways

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For the inhibition of MEK, we used trametinib (# S2673; Selleckchem) and cobimetinib (#S8041; Selleckchem) at indicated concentrations. Vemurafenib (#S1267; Selleckchem) and dabrafenib (#2807; Selleckchem) inhibited BRAF at indicated concentrations. For the inhibition of the p38 pathway, we used Skepinone-L (#S7214; Selleckchem) at indicated concentrations. Activation of the p38 pathway was induced by 100 μM hydrogen peroxide (H2O2) (#H1009; Sigma-Aldrich) for 1 h.
Gutierrez-Prat N., Zuberer H.L., Mangano L., Karimaddini Z., Wolf L., Tyanova S., Wellinger L.C., Marbach D., Griesser V., Pettazzoni P., Bischoff J.R., Rohle D., Palladino C, & Vivanco I. (2022). DUSP4 protects BRAF- and NRAS-mutant melanoma from oncogene overdose through modulation of MITF. Life Science Alliance, 5(9), e202101235.
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4

Chemical Characterization of AD Compounds

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Skepinone-L was purchased from SelleckChem (S7214). The AD compounds, sorafenib, and regorafenib were synthesized as previously described; details are provided in the supplemental synthetic methods section. Final compounds were characterized by NMR on a 400 MHz Bruker Avance III HD system and a Xevo TQD UPLC-mass spectrometer (Waters).
Yu J.X., Craig A.J., Duffy M.E., Villacorta-Martin C., Miguela V., de Galarreta M.R., Scopton A.P., Silber L., Maldonado A.Y., Rialdi A., Guccione E., Lujambio A., Villanueva A, & Dar A.C. (2019). Phenotype-based screens with conformation-specific inhibitors reveal p38 gamma and delta as targets for HCC polypharmacology. Molecular cancer therapeutics, 18(9), 1506-1519.
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5

Antibody and Inhibitor Protocols for Signaling Pathway Analysis

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Total antibodies against EphA2 (#6997) and phospho-specific antibodies against EphA2 (Ser-897; #6347), RSK (Ser-380; cross-reacting with RSK2 Ser-386; #11989), RSK (Thr-573; #9346), p38 (Thr-180/Tyr-182; #4511), ERK (Thr-202/Tyr-204; #9101), and MK2 (Thr-334; #3041 for Figs. 3 and 5, C–E or #3007 for Figs 5B and 6) were purchased from Cell Signaling Technology; antibodies against total ERK (C-9), p38α (A-12), RSK1 (C-21), RSK2 (C-19), MK2 (A-11), HSP27 (F-4), α-Tubulin (B-7), and β-Actin (C-4) were from Santa Cruz Biotechnology; an antibody against FLAG (F1804) was from Merck KGaA; a phospho-antibody against HSP27 (MAB23141) was from R&D Systems. Recombinant human EGF was obtained from R&D Systems; recombinant human active GST-MK2 protein was from Carna Biosciences; a phos-tag ligand, anisomycin, NaCl, and CDDP were from Wako Pure Chemical Industries; SB203580 and TMZ were from Merck KGaA; MK2 inhibitor III and GSK2334470 were from MedChemExpress; BI-D1870 was from BioVision; BIRB796, Skepinone-L, and BX-702 were from Selleck Chemicals. All chemical inhibitors were dissolved in dimethyl sulfoxide (DMSO).
Zhou Y., Oki R., Tanaka A., Song L., Takashima A., Hamada N., Yokoyama S., Yano S, & Sakurai H. (2023). Cellular stress induces non-canonical activation of the receptor tyrosine kinase EphA2 through the p38-MK2-RSK signaling pathway. The Journal of Biological Chemistry, 299(5), 104699.
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