The A549 and H460 human NSCLC cell lines and HEK293 cells were obtained from the American Type Culture Collection (ATCC) and their identity verified by STR profiling. The murine cell lines (344SQ, 531LN2) were derived from KrasLA1/+ /Trp53R172HΔg/+ mice [38 (link), 39 (link)]. For some experiments, cells were treated with vehicle (0.01% DMSO) or the SRC inhibitor dasatinib (MedChemExpress, Monmouth Junction, NJ, 50 nM for the A549 cell line, and 500 nM for the H460 cell line), or 2-Deoxy-D-glucose (2DG; MedChemExpress, Monmouth Junction, NJ, 10 mM for all cell lines).
Dasatinib
Manufactured by MedChemExpress
Sourced in United States
Dasatinib is a tyrosine kinase inhibitor. It is a synthetic small molecule that inhibits multiple tyrosine kinases, including BCR-ABL, SRC, c-KIT, EPHA2, and PDGFRβ.
Automatically generated - may contain errors
Lab products found in correlation
Bosutinib, by MedChemExpress (2 mentions)
Trametinib, by MedChemExpress (2 mentions)
Nilotinib, by MedChemExpress (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
2 deoxy d glucose, by MedChemExpress (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Vandetanib, by MedChemExpress (1 mentions)
Lapatinib, by MedChemExpress (1 mentions)
Ibrutinib, by MedChemExpress (1 mentions)
Sunitinib, by MedChemExpress (1 mentions)
Gilteritinib, by MedChemExpress (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Hek293 cell, by American Type Culture Collection (1 mentions)
Tak 242, by MedChemExpress (1 mentions)
Gdc 0994, by MedChemExpress (1 mentions)
Bi8622, by MedChemExpress (1 mentions)
Crizotinib, by MedChemExpress (1 mentions)
Tivantinib, by MedChemExpress (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
17 protocols using dasatinib
1
NSCLC Cell Line Characterization and Treatment
2
Characterization of OCT1 Transporter Activity
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Human embryonic kidney (HEK293) cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA). HEK293 cells stably transfected with mouse OCT1 (mOCT1) or human OCT1 (hOCT1) were cultured in Dulbecco’s Modified Eagle Media (DMEM) media supplemented with 10% fetal bovine serum (FBS) and grown at 37°C in a humidified incubator containing 5% CO2. Radiolabeled [14C] TEA and [14C] metformin were obtained from American Radiochemicals (St. Louis, MO). Cellular uptake assays were performed 48 h following transient transfection by Lipofectamine 3000 Transfection Reagent (Thermo Fisher Scientific, Waltham, MA). ON-TARGETplus Human YES1 siRNA was obtained from Dharmacon (Lafayette, CO). RNA extraction kits were obtained from Omega Bio-tek (Norcross, GA). Reference standards of decynium22, a positive control inhibitor, as well as the TKIs bosutinib, dasatinib, gilteritinib, ibrutinib, lapatinib, sunitinib, vandetanib, and CH6953755 were obtained from MedChemExpress (Monmouth Junction, NJ).
3
TLR4, Caveolae and Src Signaling in E. coli Interactions
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For analysis of the interactions among TLR4, caveolae-dependent endocytosis, and Src signalling in E. coli internalization and elimination, monocytes were pretreated for 2 h with either 2 μmol/L Tak242 (MedChemExpress, Monmouth Junction, USA) —a TLR4 inhibitor, 3 μmol/L filipin—a caveolae-mediated endocytosis inhibitor (MedChemExpress), or 3 μmol/L dasatinib (MedChemExpress) —an Src inhibitor. After treatment, the cells were used for subsequent experiments.
4
Kinase Inhibitor Treatment of Jurkat Cells
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TAK-733 is a selective MEK inhibitor (Jasek-Gajda et al., 2018 (link)) and GDC0994 is an inhibitor of ERK (Kirouac et al., 2017 (link)). Jurkat cells were treated with 1 μM TAK-733 (MedChemExpress, NJ, United States) or 1 μM GDC0994 (MedChemExpress) for 0, 1, 2, 4, 6 and 8 h.
Dasatinib is an Src family kinase inhibitor and Src can phosphorylate the Y283 of E26 transformation-specific-1 (Ets-1), activate Ets-1 and increase its stability (Lu et al., 2014 (link); Appel et al., 2017 (link)). Jurkat cells were treated with 10 μM Dasatinib (AbMole, Houston, TX, United States) for 0, 1, 2, 4, 6 and 8 h.
BI8622 is a specific inhibitor of HUWE1 (Peter et al., 2014 (link)). BI8622 with an IC50 of 3.1 μM was from MedChemExpress. Jurkat cells were treated with 1 μM BI8622.
Dasatinib is an Src family kinase inhibitor and Src can phosphorylate the Y283 of E26 transformation-specific-1 (Ets-1), activate Ets-1 and increase its stability (Lu et al., 2014 (link); Appel et al., 2017 (link)). Jurkat cells were treated with 10 μM Dasatinib (AbMole, Houston, TX, United States) for 0, 1, 2, 4, 6 and 8 h.
BI8622 is a specific inhibitor of HUWE1 (Peter et al., 2014 (link)). BI8622 with an IC50 of 3.1 μM was from MedChemExpress. Jurkat cells were treated with 1 μM BI8622.
5
Investigating Lung Cancer Cell Lines
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ERL, tivantinib, dasatinib, osimertinib, olmutinib, crizotinib, and cabozantinib were purchased from MedChemExpress LLC (Princeton, NJ, USA). CDODA-Me was kindly donated by Dr. Stephen Safe (Texas A & M University, College Station, TX, USA). RPMI 1640 medium, fetal bovine serum (FBS), crystal violet, hank balanced salt solution (HBSS), and penicillin-streptomycin-neomycin were purchased from Sigma Aldrich (St. Louis, MO, USA). Primary antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA), and β-actin was purchased from Santa Cruz Biotechnology, Inc. (Carlsbad, CA, USA). The non-small cell lung cancer cell line, HCC827, was purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA); the HCC827R cell line (ERL-resistant) was provided by Dr. Arun Rishi (Wayne State University, Detroit, MI, USA).
6
Profiling NSCLC Cell Lines with Kinase Inhibitors
7
Cytotoxicity Assay of Cancer Drugs
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PC9 and PC9GR cells in logarithmic growth stage were seeded in 96-well plates at a density of 3,000 cells per well and grown overnight. The next day, the growth medium was replaced with fresh media with dasatinib (MedChemExpress, Monmouth Junction, NJ, USA), pluripotin (MedChemExpress, Monmouth Junction, NJ, USA), trametinib (MedChemExpress, Monmouth Junction, NJ, USA), and KPT-185 (MedChemExpress, Monmouth Junction, NJ, USA), respectively, by the gradient dilution method. After being incubated for 72 h, Cell Counting Kit 8 (APExBIO, Houston, Texas, USA) was added for an additional 2 h of incubation at 37°C. Cell viability was determined by measuring the absorbance at 450 nm in a microplate reader (Thermo, Waltham, MA, USA).
8
Combinatorial Anticancer Drug Screening
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A549 cells were seeded in 96-well plates at a density of 2x103/well and maintained in an incubator overnight. After the removal of the culturing medium, and the cells were fed with fresh medium containing dasatinib, selumetinib, and MK-2206 (MedChemExpress LLC. Monmouth Junction, USA) alone, or in combination. Cell viability was measured by the Dojindo cell counting kit-8 (CCK-8, GlpBio, USA) at 24, 48, and 72 hours.
9
Senescence Reversal in Ovarian Cells
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COV434 cells and primary pT&S cells were transfected with siRNA (RIBOBIO) targeting FOXP1, SOX4 and FOS using Lipofectamine 3000 (Thermo, L3000015). The sequences of the siRNAs are listed in Supplementary Table 5 . In brief, cells were incubated in 50 nM siRNAs for 6 h and cultured in DMEM containing 10% FBS for another 48 h. For exploring senolytics fighting ovarian cell senescence, cells were treated with quercetin (10 μM, MedChemExpress), dasatinib (100 nM, MedChemExpress) or fisetin (10 μM, MedChemExpress) for 24 h. After that, cells were used for RT–qPCR, western blot, SA-β-gal staining and immunofluorescence.
10
Transwell Migration Assay with Inhibitors
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Transwell experiments were performed as described previously [66 (link)]. PLC/PRF/5 or SK-Hep-1 cells under indicated treatments were deprived of serum for 24 h before analyses. A total of 1 × 105 cells were seeded to the upper chamber in transwell inserts (BD Biosciences) with serum-free medium, and medium containing 10% FBS was added into the lower chamber of 24-well plate (Corning). After additional 48 h, migrating cells adhered to the lower surface of filter were washed with PBS, fixed with 20% methanol for 20 min and stained with 0.1% crystal violet (Sangon Biotech). For SRC inhibitors treatment, cells were treated with PP2 (10 mM for 24 h, MedChemExpress) or Dasatinib (0.1 mM for 24 h, MedChemExpress) or Saracatinib (1 mM for 6 h, MedChemExpress) before seeding in chambers. The numbers of migrating cell per well were counted under a light microscope in eight predetermined fields. Data are presented as means ± SD of values from these eight fields.
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