Cell viability assessments were carried out essentially as described previously (36 (link)). Briefly, 25 nL of 1000x 1:1 serially diluted Src inhibitors (Bosutinib or Saracatinib, Selleck Chemicals) were spotted in quadruplicate onto a 384-well tissue culture treated plate (Greiner) using a Labcyte Echo acoustic dispensing device. Py230 cells were resuspended in 20 μM of Hydroxychloroquine (HCQ) or vehicle at 1x105 cells/mL, and 25 μl (2,500 cells) were plated per well. Cells were incubated at 37°C for 48 hr before CellTiterGlo (Promega) was utilized as a proxy for cell viability on a Synergy 2 plate reader using Gen5 software (BioTek, Winooski, VT). Percent viability was calculated by normalizing to vehicle only controls in Microsoft Excel and plotted using GraphPad Prism.
Bosutinib
Manufactured by Selleck Chemicals
Sourced in United States, Germany
Bosutinib is a small molecule tyrosine kinase inhibitor. It is used as a laboratory reagent for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Dasatinib, by Selleck Chemicals (16 mentions)
Saracatinib, by Selleck Chemicals (12 mentions)
Ponatinib, by Selleck Chemicals (5 mentions)
Imatinib, by Selleck Chemicals (5 mentions)
Erlotinib, by Selleck Chemicals (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Trametinib, by Selleck Chemicals (3 mentions)
Sb203580, by Selleck Chemicals (3 mentions)
Ibrutinib, by Selleck Chemicals (2 mentions)
Fedratinib, by Selleck Chemicals (2 mentions)
Selumetinib, by Selleck Chemicals (2 mentions)
Fibronectin, by Merck Group (2 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (2 mentions)
Dasatinib, by Chemietek (2 mentions)
Gefitinib, by Selleck Chemicals (2 mentions)
Sch772984, by Selleck Chemicals (2 mentions)
Tofacitinib, by Selleck Chemicals (2 mentions)
Ruxolitinib, by Selleck Chemicals (2 mentions)
Dasatinib, by Cell Guidance Systems (2 mentions)
Nilotinib, by Selleck Chemicals (2 mentions)
35 protocols using bosutinib
1
Cell Viability Assay for Src Inhibitors
2
Mass Spectrometry Analysis of Screened Compounds
3
Antibodies and Pharmacological Modulators for AMPK Signaling
4
Establishing and Characterizing NPC Cell Lines
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HONE1, HK1, CNE1, CNE2, SUNE1, C666-1, and NPC43 NPC cell lines, and immortalized nasopharyngeal epithelial cell lines NP69 and NP460 were cultured as previously described [4 (link),18 (link)]. Cell lines were authenticated by and obtained from the Hong Kong NPC AoE Cell Line Repository. HK11.19 is a HONE1/chromosome 11 microcell hybrid cell line (MCH) that was established by the transfer of an additional intact human chromosome 11 into HONE1 and expressed physiological levels of THY1 (in the recipient HONE1 cells, THY1 expression is downregulated by promoter hymermethylation, and human THY1 maps to chromosome 11q22.3) [4 (link),19 (link)]. Recombinant human PDGF-BB was obtained from PeproTech (PeproTech, Rocky Hill, NJ, USA, #100-14B-10). KX2-391 (Selleckchem, Houston, TX, USA, #S2700), bosutinib (Selleckchem, Houston, TX, USA, #S1014), and saracatinib (Selleckchem, Houston, TX, USA, #1006) were prepared in DMSO at a concentration of 10 mM for an in vitro assay. For the in vivo assay, KX2-391 was first dissolved in DMSO to a concentration of 85 mg/mL, then formulated with PEG-300 (Sigma-Aldrich, St. Louis, MI, USA, #202371) and PBS before injection following the manufacturer’s instructions (the injection solution contains 4% DMSO, 30% PEG-300, and 66% PBS).
5
Compound Preparation for Cell Viability Assays
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Enzalutamide (MCE, HY-70002, NJ, US), EPI-001 (Selleck, S7955, TX, US), R1881 (Sigma, R0908, MO, US), Afatinib (Selleck, S1011, TX, US), Bosutinib (Selleck, S1014, TX, US), PD98059 (Selleck, S1177, TX, US), Cycloheximide (MCE, HY-12320, NJ, US), and Tunicamycin (MCE, HY-A0098, NJ, US) were stored as stock solutions in DMSO (Sigma, MO, US).
6
Cellular SRC Inhibitor Assay
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The SRC inhibitors (dasatinib, bosutinib, and saracatinib) were purchased from Selleckchem (Houston, TX, USA). All chemicals were dissolved in dimethyl sulfoxide (DMSO) and aliquots were stored at −80 °C. Primary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) and horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
7
Optimized Compound Screening Protocol
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Drugs were purchased from the following: NVP-BHG712 (S2202), AZD (S7106), CCT129202 (S1519), Nintedanib (S1010), R406 (S2194), Axitinib (S1005), Bosutinib (S1014), selumetinib (S1008), Navitoclax (ABT263, S1001), PF561772 (S2890, FAK inhibitor), and SB202190 (S1077, P38 inhibitor) from Selleckchem (Houston, TX, USA). Staurosporine (S9300), etoposide (E1383), and H2O2 (216763) from Sigma-Aldrich (St. Louis, MO, USA). They were prepared 10 mM stock solution in DMSO. They were added in culture medium to obtain a suitable working solution. As a vehicle, DMSO as the same volume was added in culture medium. The stock solution was stored at -20 °C.
8
Transwell Migration Assay for MCF-7 Cells
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MCF-7 cells were starved for 6 h in DMEM with 1% FBS. Top chambers of 24-well Transwell plates (Corning, Corning, NY, USA) were pretreated with 1% matrigel (BD Biosciences, Franklin Lakes, NJ, USA) in phosphate-buffered saline and incubated for 1 h at room temperature. Transfected and untransfected cells were added to the top chambers of the plates. Medium with specific concentration of SIRT1 inhibitor Ex527 (0.2, 1, 5 µmol/L)19 (link) and Src inhibitor bosutinib (50, 250, 1,250 nmol/L)20 (link) and their combinations (both purchased from Selleck Chemicals, Houston, TX, USA) were added to both chambers. After 24 h of incubation, the cells on the top were removed and the cells on the bottom of the filter were fixed and stained with 2% crystal violet solution in ethanol (Beyotime Biotechnology, Shanghai, People’s Republic of China). The number of migrating cells was counted in 5 fields under an IX71 fluorescence microscope (Olympus, Tokyo, Japan), and the mean for each chamber was determined. Results are shown as the average of at least 3 independent experiments.
9
Protein Expression Modulation Assay
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To reduce the indicated protein expression, cells were transfected with indicated siRNAs (160 nM) or with scrambled control siRNAs and harvested 48 h later. The final concentration of fibronectin (Sigma-Aldrich) was 100 ng/ml. SRC inhibitors saracatinib, bosutinib, PP1, and FAK inhibitor FAK-14 (used at 10 µM each) were purchased from Selleckchem. After overnight incubation in serum-free media, cells were treated with indicated ligand or inhibitors for 15–120 min.
10
Phospho-protein Analysis of Leukemia Cells
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