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3 protocols using 4 6 diamidino 2 phenylindole (dapi)

1

Dual-Responsive Nanoparticle Synthesis

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MEO2MA, OEGMA, bis[2-(2′-broMoisobutyryloxy)-ethyl]disulfide (BiBOEDS), silver trifluoroacetate (CF3COOAg) and poly(vinylpyrrolidone) (PVP, Mw=55kD) were obtained from Sigma-Aldrich Corp (St Louis, MO, USA). Tris(2-dimethyl-aminoethyl)amine (Me6TREN) and anhydrous sodium hydrosulfide (NaHS) were purchased from Alfa Aesar (Shanghai, China). Ethylene glycol (EG, purity>99.0%), cuprous chloride (CuCl), chloroauric acid hydrate (HAuCl4·4H2O, Au content>47.8%) and 4'6-diamidino-2-phenylindole (DAPI) were purchased from Sinopharm Chemical Reagent Co., Ltd (Beijing, China). DMEM, RPMI 1640 and fetal bovine serum (FBS) were obtained from Gibco BRL/Life Technologies (Grand Island, NY, USA). pHLIP (ACEQNPIYWARYADWLFTTPLLLLDLALLVDADEGT) was synthesized by Bankpeptide Inc. (Heifei, China). All other reagents were of analytical grade and used without any further purification.
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2

Synthesis and Characterization of Nanoparticles

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Ethylene glycol and 4'6-diamidino-2-phenylindole (DAPI) were purchased from Sinopharm Chemical Reagent Co. Ltd (Beijing, China). NaSH, CF3COOAg, HAuCl4 and poly(vinylpyrrolidone) (PVP, average Mr=55,000) were purchased from Sigam-Aldrich Corp (St Louis, MO, USA). Doxorubicin hydrochloride (DOX·HCl, with the purity of above 98.0%) was obtained from Beijing Huafeng United Technology CO., Ltd. (Beijing, China). RPMI 1640, DMEM and fetal bovine serum (FBS) were purchased from Gibco BRL/Life Technologies (Grand Island, NY, USA). CCK-8 reagent was obtained from Dojindo Molecular Technologies, Inc. (Tokyo, Japan). Rabbit anti-mouse β-actin monoclonal antibody, rabbit anti-mouse CD47 polyclonal antibody and rabbit anti-mouse CD54 monoclonal antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). TRIzol reagent was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). PrimeScript RT reagent kit and SYBR Green Supermix were obtained from Takara Bio Inc. (Kusatsu, Japan). All other reagents were of analytical grade and used without any further purification.
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3

Oxidative Stress Analysis Protocol

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Pyrrole, 4-carbomethoxybenzaldehyde, propionic acid, MnCl2·4H2O, tetrahydrofuran (THF), ZrOCl2·8H2O, benzoic acid, DMF and 4'6-diamidino-2-phenylindole (DAPI) were purchased from Sinopharm Chemical Reagent Co. Ltd (Beijing, China). RPMI 1640 and fetal bovine serum (FBS) were purchased from Gibco RBL/Life Technologies (Grand Island, NY, USA). SOSG was obtained from Thermo Fisher Scientific Inc. (Waltham, MA, USA). 2'7'-DCFH-DA was purchased from Sigma-Aldrich Company (St Louis, MO, USA). GSH assay kit, GPX assay kit and LPO assay kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All other regents were of analytical grade and used without any further purification.
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