1 2 dimyristoyl sn glycero 3 phosphocholine dmpc

We used 10 mM Phosphate buffer, pH 7.4, adjusted with 1 M HCl or 1 M NaOH at 25 °C. 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol), sodium salt (DMPG), and cholesterol (Chol) lipids were purchased from Sigma-Aldrich (Darmstadt, Germany). In this work, amphiphilic phosphorus dendrons of the first (D1) and second (D2) generation were prepared and studied according to [37 (link)] (Figure 1). The chemical structure and molar mass of the first-generation dendron were C₁₃₈H₂₂₄C_l10N₃₄O₇P₈S₅ and Mw = 3234.13 g/mol, and the chemical structure of the second generation was C₂₆₈H₄₂₄C_l20N₇₄O₁₇P₁₈S₁₅ with a molar mass Mw = 6702.27 g/mol. Phosphate buffer, pH 7.4, purchased from Sigma-Aldrich (Darmstadt, Germany), was used to isolate the exosomes.

1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), CaCl₂, EDTA, gentamycin sulfate, MβCD, 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyl-tetrazolium bromide (MTT), MgCl₂, Na₂HPO₄, penicillin, phenylmethylsulfonyl fluoride (PMSF), sodium bicarbonate, streptomycin, and Tris were obtained from Sigma Chemical Co. (St. Louis, MO). DMEM/F-12 [Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (Ham) (1:1)] and fetal calf serum (FCS) were from Gibco/Life Technologies (Grand Island, NY). Bicinchoninic acid (BCA) reagent for protein estimation was from Pierce (Rockford, IL). Lovastatin was obtained from Calbiochem (San Diego, CA). 4-(2-(6-(Dioctylamino)-2-naphthalenyl)ethenyl)-1-(3-sulfopropyl)-pyridinium inner salt (di-8-ANEPPS) was purchased from Molecular Probes (Eugene, OR). The concentration of stock solution of di-8-ANEPPS in methanol was estimated from its molar extinction coefficient (ε) of 37,000 M⁻¹cm⁻¹ at 498 nm^{46 (link)}. All other chemicals used were of the highest available purity. Water was purified through a Millipore (Bedford, MA) Milli-Q system and used throughout.

Naringenin (98%), product no. W530098; quercetin (≥95%), product no. Q4951; (+)-catechin hydrate (≥98%) product no. C1251; 1,6-diphenyl-1,3,5-hexatriene (DPH), 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) were from Merck/Sigma-Aldrich (St Louis, MO, USA, or Steinheim, Germany). Mueller–Hinton (MH) agar and Mueller–Hinton (MH) broth suitable for microbiology were supplied by Oxoid (Basingstoke, UK). Other reagents and organic solvents were of analytical grade, purchased from POCh (Gliwice, Poland) and used without further purification. Sheep blood was obtained from GrasoBIOTECH (Graso Company, Starogard Gdański, Poland). The flavonoids were used as freshly prepared stock solutions (15 mM) in ethanol. In the preliminary experiments, we showed that ethanol, at the concentrations used (not exceeding 1.3%), did not considerably affect the parameters measured.