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3 protocols using poly d g c 2 p9389

1

Synthesis and Characterization of Naphthalene-based Ligand

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CuCl2, 2,7-bis(bromomethyl)naphthalene, sodium sulphate and potassium carbonate were purchased from TCI Europe and used without further purification. Bis(2-pyridylmethyl)amine was synthesized as previously reported (35 (link)). Deuterated chloroform was obtained from Apollo Scientific. 1H NMR spectra were recorded on Jeol ECX-400 and Varian 500 AR spectrometers at room temperature and all chemical shifts are relative to the residual solvent peak. ESI Mass Spectra were recorded on a Waters LCT Premier XE Spectrometer in positive mode. Microanalysis (C, H and N) was carried out using a Perkin Elmer 2400 series II analyzer. UV–Vis spectra were recorded on a Jasco UV–Vis spectrophotometer in phosphate buffer (pH 6.8). Emission experiments were conducted on a Shimadzu RF-5301 spectrofluorometer at room temperature. Thermal melting analysis was conducted on an Agilent Cary 100 dual beam spectrophotometer equipped with a 6 × 6 Peltier multicell system with temperature controller. Circular dichroism spectra were recorded on an Applied PhotoPhysics Chirascan Plus. Fluorescence spectra were recorded on a Perkin Elmer LC55. Calf thymus DNA was purchased from Invitrogen (15633019) while DNA from salmon testes (D1626) and synthetic alternating co-polymers (poly[d(A-T)2] (P0883) and poly[d(G-C)2] (P9389)) were obtained from Sigma Aldrich Ireland.
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2

Comprehensive Reagent Sourcing for Complex Syntheses

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All chemicals used for complex synthesis were purchased from Sigma Aldrich without further purification. The following assays were purchased from Merck Millipore and procedures were followed as per manufacturer protocols: Guava Nexin® Reagent (4500-0450), Guava EasyCyte™ MitoPotential Kit (4500-0250), Guava Caspase 8 FAM and Caspase 9 SR (4500-0640) and Guava Caspase 3/7. Propidium iodide (PI, BTIU40017) was purchased from VWR. RNase A (12091-021), Alexa Flour 488 goat anti-mouse IgG F(ab)2 fragment (A-11020), Alexa flour 488-phalloidin (A12379), DAPI (D1306) and Mitotracker Deep Red (M22426) were purchased from Biosciences Ireland. Anti-phospho-histone H2AX (05−636) was purchased from Merck Millipore. Salmon testes DNA (D1626) and synthetic double stranded alternating co-polymers, Poly[d(G-C)2] (P9389) and Poly[d(A-T)2] (P0883) used in CD studies were purchased from Sigma Aldrich. pUC19 plasmid DNA (N3041), CutSmart® buffer (B7204), 100X BSA (B9000) and topoisomerase I (E. coli) (M0301) were all purchased from New England Biolabs. LC3 isoform LC3A rabbit monoclonal antibody (Cell Signalling) was kindly donated by Dr. Joanne Keenan while goat anti-rabbit conjugated Alex Fluor-647 (ThermoFisher) was donated by Dr. Clair Gallagher.
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3

Comprehensive Biomolecular Characterization Assays

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All chemicals used for complex synthesis were purchased from Sigma-Aldrich without further purification. The following assays were purchased from Merck Millipore and procedures were followed as per the manufacturer protocol: Guava Nexin® Reagent (4500-0450), Guava EasyCyte™ MitoPotential Kit (4500-0250), Guava Caspase 8 6-carboxyfluorescein (FAM) and Caspase 9 SR (4500-0640) and Guava Caspase 3/7 (4500-0540). Propidium iodide (BTIU40017) was purchased from VWR. Carbonyl cyanide m-chlorophenylhydrazone (10175140), calf thymus DNA (15633-019), RNase A (12091-021), Alexa Flour 488 goat anti-mouse IgG F(ab)2 fragment (A11020), Alexa flour 488-phalloidin (A12379), 4′,6-diamidino-2-phenylindole (DAPI; D1306), NucBlue Live (R37605), singlet oxygen sensor green (S36002) and MitoTracker Deep Red (M22426) were purchased from ThermoFisher Scientific. Anti-phospho-histone H2AX (05-636) was purchased from Merck Millipore. Doxorubicin (Dox) hydrochloride (D2975000), dihydroethidium (D7008), salmon testes DNA (D1626), synthetic double stranded alternating co-polymers, poly[d(G⋅C)2] (P9389) and poly[d(A⋅T)2] (P0883) and Micrococcus Lysodeikticus (D8259) were purchased from Sigma-Aldrich. pUC19 plasmid DNA (N3041), CutSmart® buffer (B7204), 100× bovine serum albumin (BSA) (B9000) and topoisomerase I (E. coli) (M0301) were all purchased from New England Biolabs.
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