Figure 3
WB analysis of a representative NDE purification from the serum of healthy donors. (a) Quality analysis of extracellular vesicle preparation. Extracellular vesicles samples are not contaminated by Golgi apparatus (GM130), endoplasmic reticulum (Erp57), lysosomes (CD107) and nucleus (lamin A/C). The presence of the HDL marker APO A1 is detectable in both total extracellular vesicles (T) and total extracellular vesicles depleted to NDE (T-N), but disappear in NDEs fractions (N). CD9 is a common exosome marker. (b) NDE characterization. WB analysis show an enrichment of neuronal markers (NSE and SV-2a) in NDEs with respect to the other two extracellular vesicles fractions. Moreover, NDE samples are negative to the astrocytes and the post synaptic markers GFAP and PSD95, respectively. (c) STX-1a and SNAP-25 are enriched in NDE fractions. The levels of both SNARE proteins are increased in NDE with respect to the two other extracellular vesicles fractions. (d) Representative EM analysis of NDE. The ultrastructural analysis of NDE shows the presence of rounded-shaped vesicles with a diameter range of 70–100 nm. Bars correspond to 50 nm. For WB analysis 100 µg of whole PBMCs, 50 µg of serum (SE), extracellular vesicles fractions (T, N, T-N) and 0.5 µg of mouse brain cortex lysate have been loaded in each lane. Uncropped WB in (a)–(c) have been reported in Fig. S6 online.
