Figure 2

Preparation of AMP delivery system using AuNP-Apt^His conjugates (A) Schematic diagram illustrating the process of His-tagged HPA3P (HPA3P^His) loaded on His-aptamer-functionalized AuNP (AuNP-Apt^His) and the delivery of HPA3P^His to Vibrio vulnificus-infected host. (B) The binding capacity of AuNP-Apt^His to HPA3P^His was determined. The indicated concentrations of HPA3P^His were incubated with AuNP-Apt^His (1 nM), and AuNP-Apt^His-bound HPA3P^His (bound HPA3P^His) were analyzed by Tricine-SDS-PAGE and immunoblotting using His antibody. The amount of HPA3P^His bound to AuNP-Apt^His was analyzed by quantification of band intensities and compared with the standards where known amounts of HPA3P^His (total HPA3P^His) were electrophoresed. The dissociation constant (K _D) was obtained from the slope of the graphs presented. (C) The size distribution of AuNP-Apt^His and AuNP-Apt^His-HPA3P^His analyzed by dynamic light scattering (DLS) is shown. (D) Representative confocal immunofluorescence images of HeLa cells treated with buffer, AuNP-Apt^His, HPA3P^His, or AuNP-Apt^His-HPA3P^His for 10 min are presented. The cells were stained with rabbit anti-His followed by rabbit IgG-labeled with Alexa 488 (green), and the nuclei were stained with DAPI (blue). Scale bar = 20 μm.