Fig. 2

Characterization of exosomes purified from the serum of CRPS patients. Exosomes were isolated from the serum of CRPS patients using differential ultracentrifugation. The exosomes were observed by TEM under a Toshiba H8000 or JEM1230 microscope. Exosomes were fixed and negatively stained with uranyl acetate for morphology (a, b) or immunolabelled for the exosome marker CD81 and probed with 6 nm gold secondary antibody, following negative staining (c). Exosome proteins isolated from PE patient serum (Exo_s) were resolved on a reducing 12% SDS-PAGE and western blotting was used to confirm the presence of exosome marker protein CD63 (d). Nanoparticle tracking analysis of serum exosomes indicated particles with a mean diameter of 85.7 ± 0.9 nm and a particle concentration of 3.94 × 10¹¹ ± 1.6 × 10¹⁰ particles/ml (e)