Fig. 5
Generation and characterization of peptide-loaded SyEV. a Structure of fluorophore and cholesterol-conjugated peptides targeting Myd88 signaling molecules. b Flowchart of the SyEV loading procedure with the peptides. Co-incubation of MSC membranes and peptides via sonication resulted in the peptide-loaded SyEV (SyEVMyd88) that were then purified by iodixanol-based density gradient ultracentrifugation. c The characteristics of non-loaded and peptide-loaded SyEV, including the loading efficiency and vesicular size. d The loading efficiency of peptides in high pH-untreated NV/EV or treated SyEV (n = 3). e Morphology of SyEVMyd88 analyzed by TEM. Scale bars, 200 nm. f Optimization of conditions for loading of SyEV with peptides. The quantity of peptides in the final samples following ultracentrifugation was assessed by the fluorescent signal of samples containing SyEV (+ SyEV) or without SyEV (–SyEV) at various incubation temperatures (n = 3). Data are presented as the mean ± SEM. ***P < 0.001 by one-way ANOVA with Tukey’s post test