Gefitinib

1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide (polyethylene glycol)-2000] (DSPE-PEG2000-Mal), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-carboxyfluorescein (CFPE) were bought from Avanti Polar Lipids (Alabaster, AL). Phycoerythrin-labeled CD133 antibodies and the CD133 MicroBead Kit were bought from Miltenyi Biotec (Auburn, CA). Gefitinib was bought from Dalian Meilun Biotech (Dalian, China). The primers and thiolated CD133 aptamer with a sequence of 5′-SH-CCCUCCUACAUAGGG-3′ were bought from Ruibo Co., Ltd. (Guangzhou, China). N-Hydroxysuccinimide (NHS), 1-ethyl-3-3-dimethylaminopropyl carbodiimide (EDC), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were bought from Sigma-Aldrich (St. Louis, MO). The StemPro® Accutase® Cell Dissociation Reagent, the Reverse Transcription System kit, TRIzol® Reagent, B27, and insulin-transferrin-selenium (ITS) were bought from Thermo Fisher Scientific Inc. All organic reagents (analytical grade) were provided by Sinopharm (Shanghai, China) unless otherwise stated.

Brucea javanica seeds were purchased from Guangxi Zhuang Autonomous Region, People’s Republic of China, and authenticated by Prof. Guo-Yue Zhong (Jiangxi University of Chinese Medicine). BD and BH were extracted from the seeds of B. javanica and identified on the basis of nuclear magnetic resonance (NMR) data (the separation method is described in detail in the Supplementary Material); their purity was ≥95% based on high-performance liquid chromatography (HPLC) analysis. Gefitinib was provided by Meilun (ZD1839, Dalian, China). The primary antibodies NOTCH3 (mouse), β-catenin (mouse), EGFR, phospho-EGFR (Tyr1068), FOXO3a (mouse), CBP (Lys1535)/p300 (Lys1499), and β-actin were from Cell Signaling Technology (CST, Danvers, MA, USA). Bax, Bcl-2, caspase-3, and cleaved caspase-3 were provided by Santa Cruz Biotechnology (SCBT, Dallas, TX, USA). The secondary antibodies were HRP-conjugated anti-rabbit IgG and anti-mouse IgG (Abcam, Cambridge, UK).

Cantharidin, Gefitinib, Afatinib and AG1478 were obtained from Meilun Biotech Co., Ltd. (Dalian, China). CCCP was acquired from Solarbio Science & Technology Co., Ltd. (Beijing, China). Dual Luciferase Reporter Assay Kit, miRNA primer and Lipofectamine®3000 were from Vazyme Biotech Co. Ltd. (Nanjing, China). MiR-607 mimic/inhibitor were purchased from General Biol Co., Ltd (Anhui, China). Mir-X miRNA qRT-PCR SYBR Kit and luciferase vector GV306 were bought from Genechem. Co. Ltd. (Shanghai, China). PrimeScript RT Master Mis Perfect kit and SYBR® Premix Ex TaqTM II was obtained from Takara Biomedical Technology Co., Ltd. (Beijing, China). EGFR, p53, Bax, Bcl-2, Mcl-1, mTOR, APAF-1, Caspase9, Cleaved-caspase9, AIF, Caspase3, Cleaved-caspase3, PARP1, GAPDH and Goat anti rabbit IgG were from Protein Technology Group (Chicago, Illinois, USA). p-EGFR, p-AKT, AKT, p-mTOR, p-MEK, MEK, p-ERK, ERK, PI3 Kinase Antibody Sampler Kit (including PI3K-110α, PI3K-110β, PI3K-110γ, p-PI3K-P85/P55, PI3K-P85) and Cleaved-PARP were purchased from Cell Signaling Technology, Inc. (Danvers, Massachusetts, USA). The PE Annexin V Apoptosis Detection Kit I was obtained from BD Biosciences (Franklin lake, New Jersey, USA). FITC-actin and Cy3-goat anti rabbit IgG was purchased from Yeasen Biotechnology.