Compounds were purchased from SelleckChem: RG7112 (#S7030), MG132 (#S2619), ABT-263/navitoclax (#S1001), MLN4924 (#S7109), VH298 (#S8449), and Doxorubicin (#S1208) or Sigma (Nutlin-3 #N6287 and Etoposide #E1383). YX-2–23 (RG7112 derivative) and VHL-Amine were synthesized (see Supplementary Methods for details).
Abt 263 navitoclax
Manufactured by Selleck Chemicals
Sourced in United States, Germany
ABT-263 (Navitoclax) is a laboratory research compound developed by Selleck Chemicals. It functions as a selective inhibitor of the anti-apoptotic Bcl-2 family proteins.
Automatically generated - may contain errors
Lab products found in correlation
Venetoclax abt 199, by Selleck Chemicals (5 mentions)
Propidium iodide, by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
A 1210477, by Selleck Chemicals (2 mentions)
Rpmi 1640 aqmedia, by Merck Group (2 mentions)
Abt 199 venetoclax, by Abcam (2 mentions)
Birinapant, by Selleck Chemicals (2 mentions)
S63845, by Apexbio (2 mentions)
S63845, by Selleck Chemicals (2 mentions)
Doxorubicin, by Selleck Chemicals (1 mentions)
Nutlin 3, by Selleck Chemicals (1 mentions)
Etoposide, by Selleck Chemicals (1 mentions)
Mln4924, by Selleck Chemicals (1 mentions)
Rg7112, by Selleck Chemicals (1 mentions)
Mg132, by Selleck Chemicals (1 mentions)
Recombinant mouse il 3, by Thermo Fisher Scientific (1 mentions)
Pancoll human, by PAN Biotech (1 mentions)
Easysep human basophil enrichment kit, by STEMCELL (1 mentions)
Eukitt, by Merck Group (1 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
16 protocols using abt 263 navitoclax
1
Synthesis and Evaluation of Novel Anticancer Compounds
2
Murine Myeloid Cell Culture Protocols
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C57BL/6 mice were maintained under pathogen-free conditions in IVC cages. Animal experiments were approved by the animal experimentation review board of the canton of Bern (BE31/11 and BE12/14).
RPMI-1640 AQmedia, propidium iodide (PI) and Eukitt were purchased from Sigma-Aldrich Chemie GmbH (Buchs, Switzerland). Fetal calf serum (FCS, Sera Pro, ultra-low endotoxin) was purchased from Pan Biotech (Aidenbach, Germany). 2-mercaptoethanol (2-ME), 4-hydroxytamoxifen (4-OHT), and penicillin/streptomycin were from Life Technologies (Carlsbad, CA, USA). WEHI-3B cell-conditioned medium was used as a source of mouse interleukin-3 and was produced as previously described.27 (link), 50 (link) Recombinant mouse IL-3 was purchased from Peprotech (London, UK). Q-VD-OPh was purchased from SM Biochemicals (Anaheim, CA, USA). Recombinant His6-tagged GFP-Annexin V was purified in-house as previously described.51 (link), 52 (link) Pancoll human (density 1.077 g/ml) was purchased from Pan Biotech, while EasySep Human Basophil Enrichment Kit was bought from StemCell Technologies (Vancouver, Canada). ABT-199 (Venetoclax) was manufactured by BioVision (Milpitas, CA, USA), ABT-263 (Navitoclax) and A-1210477 by Selleck Chemicals (Houston, TX, USA), while WEHI-539 hydrochloride was purchased from Hycultec (Beutelsbach, Germany).
RPMI-1640 AQmedia, propidium iodide (PI) and Eukitt were purchased from Sigma-Aldrich Chemie GmbH (Buchs, Switzerland). Fetal calf serum (FCS, Sera Pro, ultra-low endotoxin) was purchased from Pan Biotech (Aidenbach, Germany). 2-mercaptoethanol (2-ME), 4-hydroxytamoxifen (4-OHT), and penicillin/streptomycin were from Life Technologies (Carlsbad, CA, USA). WEHI-3B cell-conditioned medium was used as a source of mouse interleukin-3 and was produced as previously described.27 (link), 50 (link) Recombinant mouse IL-3 was purchased from Peprotech (London, UK). Q-VD-OPh was purchased from SM Biochemicals (Anaheim, CA, USA). Recombinant His6-tagged GFP-Annexin V was purified in-house as previously described.51 (link), 52 (link) Pancoll human (density 1.077 g/ml) was purchased from Pan Biotech, while EasySep Human Basophil Enrichment Kit was bought from StemCell Technologies (Vancouver, Canada). ABT-199 (Venetoclax) was manufactured by BioVision (Milpitas, CA, USA), ABT-263 (Navitoclax) and A-1210477 by Selleck Chemicals (Houston, TX, USA), while WEHI-539 hydrochloride was purchased from Hycultec (Beutelsbach, Germany).
3
Breast Cancer Cell Line Characterization
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T47D, MDA-MB-231, MDA-MB-468, and BT-20 cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and the Korea Cell Line Bank (KCLB, Korea). The T47D and BT20 cells were cultured in RPMI (Gibco, CA, USA) containing 10% fetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA) and 1% penicillin/streptomycin (PS; Gibco). MDA-MB-231 and MDA-MB-468 cells were cultured in DMEM (Gibco) containing 10% FBS and 1% PS. Cells were maintained at 37 °C in a humidified atmosphere of 95% air and 5% CO2 and periodically screened for Mycoplasma contamination. Both cells were confirmed by short tandem repeats (STR) DNA profiling tests in the Korean Cell Line Bank (KCLB). Caspase inhibitor z-VAD.fmk was purchased from R&D Systems, Inc. (Minneapolis, MN, USA), and SMAC mimetic LCL161 was purchased from Cayman Chemical (Ann Arbor, MI, USA). GSK’872 and necrosulfonamide (NSA) were purchased from Tocris Bioscience (Bristol, UK). ABT-263 (navitoclax) and ABT-199 (venetoclax) were obtained from Selleckchem (Houston, TX). Necrostatin-1 (Nec-1), docetaxel (DTX), Adriamycin (ADR), and cyclophosphamide (CPM) were purchased from Sigma-Aldrich (St. Louis, MO).
4
Induction and Inhibition of Cell Death Pathways
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Recombinant human TNFα was purchased from Gibco/Life Technologies (catalog no. PHC3011). TNFα-neutralizing antibody was purchased from Cell Signaling (catalog no. 7321). Birinapant was purchased from Selleckchem (catalog no. S7015). ABT263 (Navitoclax) was purchased from Selleckchem (catalog no. S1001). Z-VAD-FMK was purchased from ApexBio (catalog no. A1902). Necrostatin-1 was purchased from BioVision (catalog no. 2263-1). FasL was purchased from Enzo Life Sciences (catalog no. ALX-522-001-C010).
5
Cytotoxic Treatments for Renal Cell Carcinoma
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RCC cell lines and HEK293T cells were grown at 37 °C in an atmosphere containing 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% foetal bovine serum (FCS), 2 mM glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin. Transduced RCC cells were maintained in the medium described above supplemented with 2 μg/ml puromycin. For treatment, cells were exposed to the following substances dissolved in culture medium for defined periods of time: 0.1 to 10 μg/ml topotecan (Hycamtin®, GSK, Buehl, Germany), 100 ng/ml superkillerTrail (EnzoLifeScience, Lörrach, Germany) or 10 or 20 μM ABT263 (Navitoclax®, Selleckchem, Texas, USA) or 50 μM UO126 (Selleckchem). The corresponding negative controls were prepared with the appropriate solvent (PBS or DMSO).
6
Senolytic ABT263/Navitoclax Treatment
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On the last day before analysis, cells were treated with the senolytic ABT263/Navitoclax (Selleck Chemicals, S1001) at different concentrations for 24 h, while only vehicle (DMSO) was given as mock treatment.
7
Activation of Apoptotic Pathways in Mice
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C57BL/6J mice were maintained under pathogen-free conditions in individually ventilated cages (IVC). Animal experiments were approved by the animal experimentation review board of the canton of Bern (BE12/14, BE138/16).
RPMI-1640 AQmedia™, propidium iodide (PI), 4-hydroxytamoxifen (4-OHT), and SMI-4a were purchased from Sigma-Aldrich Chemie GmbH (Buchs, CH). Fetal calf serum (FCS, Sera Pro, ultra-low endotoxin) was purchased from Pan Biotech (Aidenbach, DE), penicillin–streptomycin from Gibco (Thermo Fisher Scientific, Waltham, MA, US). As a source of mouse IL-3, WEHI-3B cell-conditioned medium was used and produced as previously described21 (link),48 (link). Recombinant mouse IL-4, IL-3, IL-33, and IL-13 were purchased from Peprotech (London, UK). Q-VD-OPh was purchased from SM Biochemicals (Anaheim, CA, US). ABT-199 (Venetoclax/Venclexta) was manufactured by BioVision (Milpitas, CA, US), ABT-263 (Navitoclax) by Selleck Chemicals (Houston, TX, US), WEHI-539 hydrochloride was purchased from Hycultec (Beutelsbach, DE), S63845 from ApexBio (Houston, TX, US). Recombinant His6 (link)-tagged GFP-Annexin V was purified as previously described49 (link),50 (link). LY-294002 and rapamycin were from Enzo Life Science (Lausen, CH).
RPMI-1640 AQmedia™, propidium iodide (PI), 4-hydroxytamoxifen (4-OHT), and SMI-4a were purchased from Sigma-Aldrich Chemie GmbH (Buchs, CH). Fetal calf serum (FCS, Sera Pro, ultra-low endotoxin) was purchased from Pan Biotech (Aidenbach, DE), penicillin–streptomycin from Gibco (Thermo Fisher Scientific, Waltham, MA, US). As a source of mouse IL-3, WEHI-3B cell-conditioned medium was used and produced as previously described21 (link),48 (link). Recombinant mouse IL-4, IL-3, IL-33, and IL-13 were purchased from Peprotech (London, UK). Q-VD-OPh was purchased from SM Biochemicals (Anaheim, CA, US). ABT-199 (Venetoclax/Venclexta) was manufactured by BioVision (Milpitas, CA, US), ABT-263 (Navitoclax) by Selleck Chemicals (Houston, TX, US), WEHI-539 hydrochloride was purchased from Hycultec (Beutelsbach, DE), S63845 from ApexBio (Houston, TX, US). Recombinant His6 (link)-tagged GFP-Annexin V was purified as previously described49 (link),50 (link). LY-294002 and rapamycin were from Enzo Life Science (Lausen, CH).
8
Apoptosis Regulating Compounds Protocol
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Dulbecco's Modified Eagle's Medium (DMEM), trypsin-EDTA, penicillin-streptomycin and dimethyl sulfoxide (DMSO), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (M2128), Hoechst 33258 (B1155), Crystal Violet (C0755), and DCF (D6883) were purchased from Sigma-Aldrich (St. Louis, MO). All tissue culture-ware was from Nunc (Roskilde, Denmark). Biotin Blocking System, peroxidase substrate (DAB) and peroxidase buffer were from DAKO (Glostrup, Denmark). Aquatex was from Merck (Darmstadt, Germany). The ABC kit was from Vecstain (Burlingame, CA). Proteinase inhibitors were from Roche (Madrid, Spain). ECL western blotting substrate was from Pierce (Thermo Fisher Scientific, Rockford, IL). BCL-2 siRNA (h) (sc-29214), BCL-xL siRNA (h) (sc-43630), MCL-1 siRNA (h) (sc-35877) and scrambled controls were purchased from Santa Cruz Biotechnology (Dallas, TX). Lipofectamine2000 (11668-027), Novex Sharp Pre-stained Protein Standard (LC5800) and JC-1 (T-3168) were from Invitrogen Life Technologies (Carlsbad, CA). Sorafenib (BAY 43-9006, Nexavar) and Regorafenib (BAY 73-4506, Stivarga) are manufactured by Bayer. A-1155463, ABT-737, AT101, HA-14, ABT-263 (Navitoclax) and ABT-199 (Venetoclax) were purchased from Selleckchem (Houston, TX).
9
BH3-Mimetics Sensitivity Assay
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Complete medium RPMI +/+ consists of RPMI 1640 supplemented with 10% FCS, Penicillin, Streptomycin (1/100 each) and 10 mM HEPES. RPMI 1640 Medium, Penicillin and Streptomycin were obtained from Biochrom (Berlin, DE). Fetal calf serum (FCS, Sera Pro, ultra-low endotoxin) and HEPES were purchased from Pan Biotech (Aidenbach, DE) and Gibco®, Fischer Scientific, respectively. ABT-199 (Venetoclax) was obtained by BioVision (Milpitas, CA, US), ABT-263 (Navitoclax) by Selleck Chemicals (Houston, TX, US), WEHI-539 hydrochloride was purchased from Hycultec (Beutelsbach, DE), S63845 from ApexBio (Houston, TX, US). A stock solution (10 mM) of every type of used BH3-mimetics was prepared in DMSO.
10
Apoptosis-Inducing Compound Screening
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UMI-77 (APExBIO);8 (link) Bax-inhibiting peptide V5 (BIP-V5; Sigma Aldrich); MG132 (Enzo Life Sciences), etoposide (Cell Signaling Technologies); PAC1, Bam7, Birinapant, Lexibulin (CYT997), Nutlin-3a, YM155, A1210477, Navitoclax (ABT-263), Venetoclax (ABT-199) and Obatoclax mesylate were all purchased from Selleckchem. All inhibitors and stressors were dissolved in DMSO; control conditions were all treated with the appropriate amount of DMSO.
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