The following materials were commercially obtained: osimertinib (Chemscene, Monmouth Junction, NJ, USA), gefitinib (Wako Pure Chemical Industries, Osaka, Japan), erlotinib (Wako Pure Chemical Industries), and DMSO (Wako Pure Chemical Industries). Antibodies were obtained from the following sources: EGFR [EP38Y] (Abcam, Cambridge, UK), PARP [46D11] (Cell Signaling Technology, Beverly, MA, USA), EphB4 [D1C7N] (Cell Signaling Technology), β-actin [AC-15] (Sigma-Aldrich, Burlington, MO, USA), and Ki-67 [MIB1] (DAKO, Santa Clara, CA, USA).
Osimertinib
Manufactured by ChemScene
Sourced in United States
Osimertinib is a small-molecule tyrosine kinase inhibitor. It is used as a laboratory research tool to study the epidermal growth factor receptor (EGFR) pathway.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin a1978 antibody, by Merck Group (3 mentions)
Ym155, by ChemScene (2 mentions)
Dimethyl sulfoxide (dmso), by Fujifilm (1 mentions)
β actin ac 15, by Merck Group (1 mentions)
Erlotinib, by Fujifilm (1 mentions)
Parp 46d11, by Cell Signaling Technology (1 mentions)
Ephb4 d1c7n, by Cell Signaling Technology (1 mentions)
Egfr ep38y, by Abcam (1 mentions)
Gefitinib, by Fujifilm (1 mentions)
Ki 67 mib 1, by Agilent Technologies (1 mentions)
Anti egfr 4267, by Cell Signaling Technology (1 mentions)
Brexpiprazole, by Cayman Chemical (1 mentions)
Erlotinib, by ChemScene (1 mentions)
Afatinib, by ChemScene (1 mentions)
Selumetinib, by Selleck Chemicals (1 mentions)
Tranilast, by Tokyo Chemical Industry (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
Anti cleaved caspase 3 9661, by Cell Signaling Technology (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Anti lc3 3868, by Cell Signaling Technology (1 mentions)
7 protocols using osimertinib
1
EGFR Signaling Pathway Inhibition
2
Antibody and Drug Reagent Procurement
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Anti-survivin (#2808), anti-p-AKT (Ser473 #9271, Thr308 #9275), anti-AKT (#9272), anti-p-mTOR (#2974), anti-mTOR (#2972), anti-p-ERK (#9101), anti-ERK (#4695), and anti-EGFR (#4267) antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Anti-β-actin (A1978) antibody was from Sigma (St. Louis, MO, USA). Osimertinib and YM155 were purchased from Chemscene LLC. (Monmouth Junction, NJ, USA), and dissolved in dimethyl sulfoxide (DMSO) to 10 mM and 20 μM, respectively, as stock solutions. Brexpiprazole was from Cayman Chemical Company (Ann Arbor, MI, USA) and was dissolved in DMSO to 10 mM as a stock solution.
3
Synthesis and Evaluation of TAS6417
4
Investigating EGFR and KRAS NSCLC Cell Lines
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Three EGFR‐mutant NSCLC cell lines (HCC827 [EGFR exon19del E746‐A750], HCC4011 [EGFR L858R], and PC9 [EGFR exon19del E746‐A750]) and two KRAS‐mutant NSCLC cell lines (H23 [KRAS G12C] H2009 [KRAS G12A]) were used in this study. All the NSCLC cell lines were purchased from the American Type Culture Collection. Three CAFs named CAF1, CAF2, and CAF3 were obtained from surgically resected three NSCLC specimens derived from each three patient, immortalized by retroviral transduction of hTERT to stabilize the cell phenotype and avoid cellular senescence, and cultured in Roswell Park Memorial Institute (RPMI)‐1640 medium supplemented with 10% fetal bovine serum at 37°C in a humidified incubator under 5% CO2. Studies using clinical specimens were approved by the Okayama Medical School and Hospital's Research Ethics Committee (approval number: #1906‐033), and informed consent was obtained from individual patients for the use of their materials. Osimertinib, an EGFR‐tyrosine kinase inhibitor, was purchased from ChemScene; selumetinib, an MEK inhibitor, was purchased from Selleckchem; and tranilast was purchased from Tokyo Chemical Industry.
5
Autophagy Regulation in Cancer Cells
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Anti-LC3 (#3868) and anti-cleaved caspase 3 (#9661) antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). The anti-SQSTM1 (p62, sc-28359) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-β-actin (A1978) antibody was from Sigma (St. Louis, MO, USA). Osimertinib was purchased from Chemscene LLC. (Monmouth Junction, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) to 10 mM as a stock solution. Doxazosin was from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan) and dissolved in DMSO to prepare a 10 mM stock solution. 3-Methyladenine (3-MA) was from Merck Millipore (Darmstadt, Germany) and dissolved in DMSO to 75 mM as a stock solution. Bafilomycin A1 was from Sigma and dissolved in DMSO to 100 µM as a stock solution.
6
Antibody and Chemical Reagent Preparation
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An anti-survivin (#2808) antibody was purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). An anti-β-actin (A1978) antibody was from Sigma (St. Louis, MO, USA). Gemcitabine was also from Sigma, and was dissolved in dimethyl sulfoxide (DMSO) to prepare a 1 mM stock solution. Spironolactone and eplerenone were from Tokyo Kasei Kogyo (Tokyo, Japan) and dissolved in DMSO to prepare a 100 mM stock solution. Osimertinib and YM155 were purchased from Chemscene LLC. (Monmouth Junction, NJ, USA) and dissolved in DMSO to 10 mM and 20 μM, respectively, as stock solutions. Aldosterone was from Toronto Research (North York, Canada) and dissolved in DMSO to a 100 mM stock solution. Esaxerenone was from Medchem (Monmouth Junction, NJ, USA) and dissolved in DMSO to a 50 mM stock solution. MG132 was from Wako Pure Chemical Industries (Osaka, Japan) and dissolved in DMSO to a 10 mM stock solution.
7
EGFR and KRAS Mutant NSCLC Cell Lines
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Two EGFR-mutant NSCLC cell lines: HCC827 (EGFR exon19del E746-A750), H1975 (EGFR L858R and T790M), one KRAS mutant NSCLC cell line, A549 (KRAS G12S) and one pancreatic cancer cell line, PANC-1 (KRAS G12D), were used in this study. All cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA). These cell lines were cultured in Roswell Park
Memorial Institute-1640 medium supplemented with 10% fetal bovine serum at 37 ºC in humidified incubator under 5% CO2 gas. Osimertinib, EGFR-tyrosine kinase inhibitor, was purchased from ChemScene (Monmouth Junction, NJ, USA)
and monensin was purchased from Cayman Chemical (Ann Arbor, MI, USA).
TGF-β was purchased from Wako (Osaka, Japan).
Memorial Institute-1640 medium supplemented with 10% fetal bovine serum at 37 ºC in humidified incubator under 5% CO2 gas. Osimertinib, EGFR-tyrosine kinase inhibitor, was purchased from ChemScene (Monmouth Junction, NJ, USA)
and monensin was purchased from Cayman Chemical (Ann Arbor, MI, USA).
TGF-β was purchased from Wako (Osaka, Japan).
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