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2 protocols using 1 2 dioleoyl 3 trimethylammonium propane dotap

1

Synthesis of Multifunctional Nanoparticles

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Gd(CH3CO2)3xH2O (99.9%), Nd(CH3CO2)3xH2O (99.9%), Y(CH3CO2)3•4H2O (99.99%), FeCl3·6H2O (97%), citric acid hydrate, and sodium dodecylbenzenesulfonate (SDBS, 95%) were purchased from J&K Scientific Ltd. Methotrexate (MTX, 98%) and propidium iodide (PI, 94%) were purchased from Shanghai Energy Chemical Co. Ltd. Pyrrole (99%) and dithiodiglycolic acid (96%) was purchased from Tokyo Japan Chemical Industry (TCI) Co. Ltd. NH4F (98%), NaOH (97%), 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride (EDC, 97%), N‐hydroxysuccinimide sodium salt (sulfo‐NHS, 98%), and dimethyl sulfoxide (DMSO, 99.9%) were purchased from Sinopharm Chemical Reagent Co. China. 1‐Octadecene (95%), oleic acid (>95%), TNF‐α siRNA, 1,2‐dioleoyl‐3‐trimethylammonium‐propane (DOTAP), Hoechst 33 258 (98%), and Annexin V‐FITC apoptosis detection kit were purchased from Sigma‐Aldrich. All chemical reagents were purchased from commercial sources and used directly without further purification. Deionized water was used throughout.
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2

Angiopep-2 Functionalized Nanoparticles

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DSPE-PEG-2000 was obtained from Meilun Biotechnology Co., Ltd (Dalian, China). P(TMZ)n was synthesized from Beixin Biotechnology Co., Ltd (Suzhou, China). 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) and 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). AnnexinV-fluorescein isothiocyanate (FITC)/propidium iodide (PI) was obtained from Nanjing Key GEN BioTECH Co. Ltd (Nanjing, China). Calcein/PI Live/Dead Viability/Cytotoxicity Assay Kit were purchased from Beyotime Biotechnology. Angiopep-2 (TFFYGGSRGKRNNF KTEEY) was purchased from GLBiochem Ltd (Shanghai, China). Cell Counting Kit-8 (CCK8) and D-Luciferin potassium were purchased from Meilun Biotechnology Co., Ltd (Dalian, China). LysoTracker™ green was brought from Invitrogen (Carlsbad, CA, USA). MGMT Rabbit monoclonal antibody (mAb) was got from Cell Signaling Technology Co., Ltd (Danvers, MA, USA). Beta-actin mAb was bought from Protein tech Antibodies People Trust (Chicago, IL, USA). Cy5-labeled siRNA (Cy5-siRNA), negative control siRNA with a scrambled sequence (nonsense, antisense strand,5′-UCGAAGUACUCAGCGUAAGdTdT-3′), and siRNA targeting MGMT mRNA (siMGMT, antisense strand, 5′-UGUCGCUCAAACAUCCAUCdTdT-3′) were purchased from Beixin Biotechnology Co., Ltd (Suzhou, China).
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