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. 2015 Dec 30;291(17):8931–8950. doi: 10.1074/jbc.M115.695304

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FIGURE 1. — IL-15:IL-15Rα is bound to the surface of fabricated PLGA nanoparticles. a, biotinylated IL-15:IL-15Rα was incubated with avidin-coated nanoparticles to create IL-15:IL-15Rα-coated nanoparticle constructs (IL-15:IL-15Rα/NP). IL-15:IL-15Rα binding was analyzed by flow cytometry after staining with anti-human IL-15 PE. The conditions tested were IL-15:IL-15Rα/NP (blue curve), bare nanoparticles (solid gray curve), and nanoparticles blocked with excess free biotin before IL-15:IL-15Rα binding (red curve). b, TEM images of nanoparticles. IL-15:IL-15Rα/NP were stained with anti-human IL-15 and 6 nm gold-linked anti-mouse IgG1 and then imaged by TEM. Arrows indicate regions where gold nanoparticles are associated with IL-15 clusters on the surface of nanoparticles. c, TEM images of NP-treated DCs. DCs were treated with IL-15:IL-15Rα/NP or uncoated NPs and then fixed overnight in paraformaldehyde and sectioned in liquid nitrogen. Sections were stained with anti-human IL-15 and 6 nm gold-linked anti-mouse IgG1 and then imaged by TEM. Arrows indicate IL-15 labeling with gold nanoparticle-linked antibodies.