2 4 2 hydroxyethyl piperazin 1 yl ethanesulfonic acid hepes

Poly(L-lysine hydrobromide) (PLL-HBr, M_w = 7.5 × 10³) and dextran (Dex, M_w = 8.0 × 10³–1.2 × 10⁴) were obtained from Sigma-Aldrich (St. Louis, MO, USA) and Funakoshi Co. (Tokyo, Japan), respectively. Sodium hydroxide, sodium chloride and manganese(II) chloride tetrahydrate were purchased from Wako Pure Chemical Industries (Osaka, Japan). 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) was obtained from Nacalai Tesque, Inc. (Kyoto, Japan). Poly(L-lysine)-graft-Dextran (PLL-g-Dex) cationic comb-type copolymer was synthesized by a reductive amination reaction of dextran with PLL according to [14 (link)]. The resulting copolymer was purified by an ion exchange column and dialysis, and obtained by freeze drying. ¹H nuclear magnetic resonance spectrometer and gel permeation chromatography equipped with a multi angle light scattering detector were employed to characterize the resulting copolymer. PLL-g-Dex copolymer consisting of 10 wt% PLL and 90 wt% dextran (11.5 mol.% of lysine units of PLL were substituted with dextran) was used in this study (Figure 1). HPLC-grade oligonucleotide with LNA modification was purchased from Gene Design Inc. (Osaka, Japan). HPLC-grade oligonucleotides with the sequences summarized in Figure 2(a) except that with LNA modification were purchased from Fasmac Co., Ltd (Kanagawa, Japan) and used without further purification.

Egg phosphatidylcholine (EPC), 1,2-dioleoyl-sn-glycerophosphocholine (PC) and poly(ethylene glycol) (average molecular weight is 2000) dimyristoyl-rac-glycerol (PEG-DMG) were purchased from NOF Corporation (Tokyo, Japan). CHEMS and PS were obtained from Avanti Polar Lipids (Alabaster, AL, USA). Cholesterol (chol) and RPMI-1640 were obtained from Merck (Burlington, MA, USA). Chloroform, 99.5% ethanol and Dulbecco’s phosphate-buffered saline without Mg²⁺ and Ca²⁺ (PBS; KCl 200 mg/L, NaCl 8000 mg/L, KH₂PO₄ 200 mg/L and Na₂HPO₄ (anhydrous) 1150 mg/L), 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) were purchased from Nacalai tesque (Kyoto, Japan). Isoflurane was obtained from Pfizer (New York, NY, USA). Fluorescent dyes, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine (DiD) and 1,1′-dioctadecyltetramethyl indotricarbocyanine iodide (DiR) was purchased from ThermoFisher Scientific (Waltham, MA, USA). Matrigel Matrix was obtained from Becton Dickinson (Franklin Lakes, NZ, USA).