Our Hard Genuine Truth About CAPNS1

Following cellular treatment with SNAs, we have estimated that Dolutegravir and ELISA data support the notion that CAPNS1 SNAs can be naturally sorted by an endosomal-exosomal pathway. We next compared the relative ability of exo-SNAs and exosome-free SNAs to enter specific cell types. A co-cultured model was utilized to monitor the selective cellular uptake of SNAs; resting cancerous PC-3 cells were co-grown with non-cancerous endothelial C166 cells expressing GFP (C166-GFP).24 The differential populations of PC-3 and C166-GFP cells can be distinguished by their ability to fluoresce, as revealed by FACS analysis (Figure 3A). Next, Cy5-labeled SNAs were synthesized to aid in the determination of cellular uptake of the nanoconjugates (Table S1). To achieve this goal, one fraction of the Cy5-SNAs was treated with PC-3 cells to allow for exosome internalization (denoted ��exo-SNA-Cy5��). As a control, another fraction of the Cy5-SNAs did not receive cellular treatment, and therefore was free of exosomes. This control was used to investigate the uptake Talazoparib order selectivity of free SNAs by both cell types (PC-3 and C166 cells). By FACS analysis, Cy5-SNAs can be internalized into 98.4% of PC-3 and 100% of C166 cells (Figure 3B) showing no selectivity in uptake by specific cell types. By contrast, Cy5-labeled exo-SNAs that were isolated from PC-3 cells showed preferential delivery (?4 fold excess) into resting PC-3 cells relative to C166-GFP cells (18% and 4.3%, respectively) (Figure 3C). Notably, the Cy5 fluorescence of cells treated with SNAs was significantly higher (?2 fold) than the values for those treated with exo-SNAs. While this can be explained by the 3 order-of-magnitude difference in concentrations of SNAs incubated with cells in the two experiments (0.35 pM, exo-SNAs; and 300 pM, free SNAs), it also reinforces our conclusion that the exosomal encasement endows SNAs with the specific ability to target cancer cells, consistent with literature reports that exosomes derived from PC-3 express surface antigens that are specific to prostate cancer cells.