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2 protocols using as1411 aptamer

1

Targeted Delivery of PTX-Loaded Nanoparticles

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PLGA, with a lactide-glycolide ratio of 50:50 and Mw of 7,000-17,000 (ester terminated), PEI (average Mw 25,000 Da), poly (vinyl alcohol) (PVA, 30-70 kDa) and Coumarin-6 were provided by Sigma Aldrich. DiR Iodide and DiO were obtained from Yeasen. PTX was purchased from J& K Chemical Technology. The Avanti mini extruder and polycarbonate porous membrane were obtained from Avanti Polar Lipids. Antibodies targeting Rb (4H1), p-AKT (D9E), AKT (40D4), E7 (F-7) and MDR1 (D-11) were bought from Cell Signaling Technology and Santa Cruz Biotechnology, respectively. AS1411 aptamer (Ap sequence: 5′‐SH‐C6‐TTGGTGGTGGTGGTTGTGGTGGTGGTGG‐3′, 28 bp) was synthetized by Sangon Bioengineering (Shanghai, China). siRNA sequences targeting E7 (sense 5′-GCGACUCAGAGGAAGAAAATT-3′, antisense 5′-UUUUCUUCCUCUGAGUCGCTT-3′) and Cy5-labeled siRNA-NC were synthetized by Shanghai GenePharma.
The human cervical cancer cell line HeLa was obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured according to the guidelines recommended by the ATCC.
Female BALB/c nude mice were obtained from Shanghai Jiao Tong University School of Medicine and raised in the SPF animal facility. All the animal experimental procedures were conducted with the approval of the Animal Experimentation Ethics Committee of Jiao Tong University.
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2

Synthesis and Characterization of Multifunctional Nanoparticles

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Raw materials including calcium nitrate tetrahydrate (Ca(NO3)2•4H2O), sodium phosphate dodecahydrate (Na3PO4.12H2O) and sodium fluoride (NaF) were purchased from Chengdu Kelong Chemical Co., Ltd., China. The gadolinium nitrate hexahydrate (Gd(NO3)3•6H2O) and terbium nitrate pentahydrate (Tb(NO3)3•5H2O) were bought from Aladdin Chemical Inc, China. The AS1411 aptamer (GGTGGTGGTGGTTGTGGTGGTGGTGG) was synthesized by Sangon Biotech, Co., Ltd., China. The doxorubicin hydrochloride (C27H29NO11·HCl, DOX•HCl) was bought from Beijing Solarbio Science & Technology Co., Ltd., China. The Dulbecco's modified Eagle's medium (DMEM), RPMI 1640 medium, foetal bovine serum (FBS) and 0.25% trypsin‐EDTA were obtained from GIBCO, Invitrogen Co., Carlsbad, USA. A Cell Counting Kit 8 (CCK‐8) was obtained from Dojindo Laboratories, Japan, and 4′,6‐Diamidino‐2‐phenylindole (DAPI) and phalloidin were purchased from Shanghai Beyotime Biotechnology Co., Ltd., China. The ultrapure water (18.2 MΩ cm−1) obtained from a purification system (Direct 8, Milli‐Q) was utilized though the whole experiments. All chemical reagents in this study were of analytical grade.
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