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7 protocols using 1 iodododecane

1

Synthesis of DMAEMA-OEGMA Copolymers

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The monomers 2-(dimethylamino)ethyl methacrylate (DMAEMA, 98%) and (oligo ethylene glycol)methacrylate (OEGMA) (average Mn = 950 g∙mol−1, 19 ethylene oxide units) were purchased from Sigma Aldrich, Greece. Both DMAEMA and OEGMA were purified using a column filled with inhibitor removers before polymerization. 2,2′-Azobis (isobutyronitrile) (AIBN), the radical initiator utilized, was purified by recrystallization from methanol. 4-Cyano-4-(dodecylsulfanylthiocarbonyl)pentanoic acid (CDP) as the CTA, methyl iodide (CH3I) 1-iodohexane (C6H13I, ≥98%), 1-iodododecane (C12H25I, 98%), 1,4-dioxane (≥99.8% pure), and tetrahydrofuran (THF, ≥99.9% pure) were obtained from Sigma Aldrich, Greece and used as received, except 1-4-dioxane, which was first dried over molecular sieves. Deuterated chloroform (CDCl3) was used as the solvent for the 1H-NMR experiments and was also obtained from Sigma Aldrich, Greece. Dialysis tubing membranes (MEMBRA-CEL®) from regenerated cellulose of MWCO 3500 and a diameter of 22 mm were purchased from SERVA, Heidelberg, Germany.
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2

Multiwalled Carbon Nanotube Functionalization

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MWNTs (diameter 12.1 nm, s.d. 3.7 nm), synthesised by CVD, were obtained from Arkema SA (Lacq-Mourenx, France). Methyl methacrylate (MMA, > 98.5%), 4-vinyl pyridine (4-VP, 95%), poly(ethylene glycol) methacrylate (PEGMA, average Mn = 530), 1-iodododecane (IDD, 98%), iodomethane (IMe, ≥ 99%), and lithium hydroxide (LiOH, 98%) were purchased from Sigma-Aldrich for MWNTs functionalisation. Before use, all chemicals were passed through a chromatographic column consisting of neutral and basic aluminium oxide powders (aluminium oxide 90 (0.063–0.200 mm), activity stage I for column chromatography, Merck Millipore, Germany) and further degassed by bubbling N2 gas for 30 minutes, in order to remove radical inhibitors and oxygen.
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3

Synthesis of Alkylated Amines

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D-glucose, tetramethylenediamine, hexamethylenediamine, iodoethane, 1-iodopropane, 1-iodododecane were purchased from Sigma-Aldrich. Hexanal was obtained from Fluka. Octanal, decanal, dodecanal, sodium borohydride and sodium cyanoborohydride were purchased from Merck. Sodium hydroxide, potassium hydroxide, hydrochloric acid, acetic acid, methanol, 2-propanol, N,N-dimethylformamide and diethyl ether were obtained from Chempur. Nitromethane was purchased from POCH. All reagents and solvents were used as obtained without further purification.
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4

Functionalization of Carbon Nanomaterials

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MWCNTs (diameter 12.1 nm, s.d. 3.7 nm), synthesised by CVD, were obtained from Arkema SA (Lacq-Mourenx, France). Carbon black nanoparticles (Printex 90, primary particle diameter 21.1 nm, s.d. 6.2 nm) were provided by Degussa-Hüls (Frankfurt, Germany). Potassium 3-(methacryloyloxy)propane-1-sulfonate (SPMAK, 98%), (3-acrylamidopropyl)trimethylammonium chloride solution (75% in H2O), methyl methacrylate (MMA, > 98.5%), 4-vinyl pyridine (4-VP, 95%), poly(ethylene glycol) methacrylate (PEGMA, average Mn = 530), 1-iodododecane (IDD, 98%), iodomethane (IMe, ≥ 99%), and lithium hydroxide (LiOH, 98%) were purchased from Sigma-Aldrich for CNM functionalisation. Before use, all liquid monomer chemicals were passed through a chromatographic column consisting of neutral and basic aluminium oxide powders (aluminium oxide 90 (0.063–0.200 mm), activity stage I for column chromatography, Merck Millipore, Germany) and further degassed by bubbling N2 gas for at least 30 minutes, in order to remove any radical inhibitors and oxygen.
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5

Synthesis of Alkyl Halide Derivatives

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Naringenin, iodomethane, iodoethane, 1-iodopentane and 1-iodododecane were purchased from Sigma-Aldrich Chemical Co. (Steinheim, Germany), hydroxylamine hydrochloride from LOBA Feinchemie GmbH (Fischamed, Austria), anhydrous sodium acetate and potassium carbonate from Chempur (Piekary Śląskie, Poland). Anhydrous solvents were prepared according to standard procedures. All organic solvents were of analytical grade.
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6

Branched PEI Nanoparticle Synthesis

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Branched PEI2k, 1-iodododecane, ferrozine, neocuproine, ascorbic acid, lactobionic acid (LAC) and dimethylsulfoxide (DMSO) were purchased from Sigma (USA). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and ammonium acetate were purchased from Aladdin (China). All chemicals were used as received. Balb/c mice were purchased from Chengdu Dashuo Biotechnology Co. Ltd. (China). RPMI-1640, phosphate buffered saline (PBS) and penicillin/streptomycin were purchased from Hyclone (USA). Fetal bovine serum was purchased from Gibco (USA). Granulocyte-macrophage colony-stimulating factor (GM-CSF) was purchased from PeproTech (USA). Lipopolysaccharide (LPS) was purchased from Hycult Biotech (Netherlands). Annexin V-FITC/PI kit and cell counting kit-8 (CCK-8) were purchased from Dojindo (Japan). 3-Methyladenine (3-MA), wortmannin and Mammalian Cell Lysis Reagent were purchased from Sigma (USA). Protease inhibitor cocktail was purchased from Roche (USA). Mouse monoclonal LC3 antibody (NB100-2220) was obtained from Novus (USA), and mouse monoclonal p62 antibody (ab56416) was purchased from Abcam (USA). Mouse monoclonal β-Actin antibody (sc-47778) and goat-anti-mouse-IgG-HRP were purchased from Santa Cruz (USA). Polyvinylidene fluoride (PVDF) membrane and enhanced chemiluminescence (ECL) kit were purchased from Bio-Rad (USA).
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7

Diblock Copolymer Synthesis with DMAEMA and OEGMA

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Monomers 2-(dimethylamino)ethyl methacrylate (DMAEMA, 98%) and (oligo ethylene glycol)methacrylate (OEGMA) (average Mn = 475 g/mol, 9 ethylene oxide units) were purchased from Sigma Aldrich, Athens, Greece. Monomers were purified by passing through a column packed with inhibitor removers before polymerization. 2,2′-Azobis(isobutyronitrile) (AIBN) was purified by recrystallization from methanol. 4-Cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanoic acid (CPD) as the CTA, 1-iodohexane (C6H13I. ≥98%), 1-iodododecane (C12H25I, 98%) 1,4-dioxane (≥99.8% pure), which was dried over molecular sieves before use, tetrahydrofuran (THF, ≥99.9% pure), and n-hexane (≥97%) were also purchased from Sigma Aldrich, Athens, Greece. Indomethacin (IND) was obtained from Fluka, Athens, Greece and used as received. Experimental details on the diblock copolymer synthesis are provided in the Supplementary Materials section.
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