Figure 4.
Target-cell analysis determines exosome-mediated molecular transfer (Application 3). (A) Percentage of HLA-A3-positive cells detected by ISx following coculture of CD14+ cells from HLA-A3-negative individual with extracellular vesicles from HLA-A3 positive individual. Data represented as mean ± SD of two separate experiments. *P < 0.05 and **P < 0.01. (B) Representative histogram showing higher colocalization of fluorescently labeled HLA-A3 with CD63 as compared to CD45, as ascertained by the IDEAS® Similarity Bright Detail Score (SBDS). (C) Representative ISx images of single cells following 2 h coculture exhibiting discrete HLA-A3 spots and with high HLA-A3/CD63 SBDS scores (left panels), contrasted with images of cells following 24 h coculture exhibiting diffuse HLA-A3 fluorescence and with low HLA-A3/CD63 SBDS scores; suggesting incorporation and diffusion of exosome-derived HLA molecules over time. (D) Representative histogram showing the normalized frequencies of HLA-A3 positive cells exhibiting a given number of HLA-A3 “spots,” as ascertained by the IDEAS® Spot Count feature, following 2 h EV coculture. (E) Summary of proportions of HLA-A3 positive cells exhibiting a given number of HLA-A3 spots, following either 2 or 24 h EV coculture. Data presented as the mean of two separate experiments.