The 12. -O-Tetradecanoylphorbol-13-acetate (TPA), midostaurin, and dimethylsulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trypan blue solution was purchased from ThermoFisher (0.4%, Thermo Fisher Scientific, Runcorn, Cheshire, UK). Liu’s stain A and B solution was purchased from Tonyar Biotech. Inc. (Taoyuan City, Taiwan). The Cell Counting Kit-8 (CCK-8) was purchased from MCE USA (MedChemExpress, Princeton, NJ, USA).
Midostaurin
Manufactured by Merck Group
Sourced in United States
Midostaurin is a laboratory product manufactured by Merck Group. It is a small molecule that inhibits certain protein kinases. The core function of Midostaurin is to serve as a research tool for investigating cellular signaling pathways in in vitro studies.
Automatically generated - may contain errors
Lab products found in correlation
Trypan blue solution, by Thermo Fisher Scientific (1 mentions)
Venetoclax, by LC Laboratories (1 mentions)
Cytarabine, by Merck Group (1 mentions)
Crenolanib, by Adooq (1 mentions)
Ponatinib, by Adooq (1 mentions)
Edelfosine, by Bio-Techne (1 mentions)
Staurosporine, by Cell Guidance Systems (1 mentions)
Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 growth medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
6 protocols using midostaurin
1
Evaluating Cell Viability Assays
2
Macrophage Reprogramming and Leukemia Cell Assay
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HD MΦs were polarized for 7 days with M-CSF and then either reprogrammed for 7 more days in medium containing GW2580 or PLX3397 and GM-CSF or kept in M-CSF-containing medium. MΦ orientation and reprogramming was confirmed by FC. Medium was then removed, and MΦs were washed with PBS. Following this, 150 × 103 NB4 or MV-4-11 were added to MΦ monolayers in PM with inhibitors (venetoclax from LC Laboratories, or midostaurin from Sigma-Aldrich). After 48 h, cells were removed from the wells, stained for AnnV and/or 7-aminoactinomycin D (7-AAD), and analyzed by FC. Experiments were repeated using MΦs from at least three different HD.
3
Evaluating Cytotoxic Drug Combinations
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We cultured cells with or without PKC412 (Midostaurin; Sigma Aldrich, St Louis, MO, USA) alone or together (co-incubation) with daunorubicin (Scandinavian Medical Service, Helsingborg, Sweden) or cytarabine (Sigma). Cells were also cultured in daunorubicin alone and cytarabine alone. Stock solutions of 10 nM were prepared in water for all drugs and stored at − 20 °C, aliquoted to avoid repeated freeze-thawing. Final concentrations of compounds and time of incubation are indicated in the figures and/or figure text.
4
Characterization of FLT3-mutant AML Cell Lines
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Midostaurin was purchased from Sigma (St. Louis, MO, USA), crenolanib, quizartinib and ponatinib were purchased from Adooq Bioscience (Irvine, CA, USA). MV4–11 cell line was a kind gift from Dr. Mark Levis (Johns Hopkins University). MOLM-14 (FLT3-ITD, D835Y) and MOLM-14 (FLT3-ITD, F691 L) [25 (link)] were kind gifts from Dr. Neil Shah (UCSF). The cells were routinely maintained in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% fetal bovine serum at 37 °C with 5% CO2.
5
Culturing Normal and Cancer Cell Lines
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Normal human fibroblasts TIG-1, AG16409 and WI38 were from the Coriell Institute Cell Repository. The non-small cell lung carcinoma cell line H1299 and A549 were obtained from the American Type Culture Collection (ATCC), while the bladder carcinoma cell line T24 was kindly provided by Dr Anne Kiltie (University of Oxford). All cell lines were cultured in DMEM (Life Technologies) supplemented with either 15% (for the fibroblast cell lines) or 10% FBS (for cancer cell lines) at 37° C in a humidified atmosphere with 5% CO2. Cells were routinely checked for mycoplasma. Recombinant human TGF-β1 (Peprotech) was used at the indicated concentration. The PLC inhibitor (edelfosine-Tocris), H2O2 (Sigma), the PDGF inhibitor (AG 1296-Millipore), staurosporine (Cell Guidance Systems), midostaurin (Sigma) and UCN-01 (Millipore) were used at the indicated concentrations.
6
Cell Proliferation Assay Reagents
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MTT and midostaurin were commercially supplied (Sigma-Aldrich, USA). 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) was obtained from Cayman Chemicals (Ann Arbor, MI, USA). Stock solutions (10 mM in dimethyl sulfoxide (DMSO)) were prepared, which do not contain more than 0.01% DMSO in culture. RPMI 1640 growth medium and all ingredients required for complete growth medium (Penicillin-streptomycin and fetal bovine serum) were obtained from Invitrogen (Paisley, UK).
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