Figure 6
Analysis of SNARE in extracellular vesicles of IS and HC subjects. (a, c, f) Representative WB analysis of the expression levels of STX-1a and of SNAP-25 in extracellular vesicles derived from sera. Representative WB analysis of total (T) extracellular vesicles (a), NDE (N) (c) and total extracellular vesicles depleted of NDE (T-N) (f) fractions. Each sample has been tested against CD9 (total exosomes marker), NSE (NDE marker), STX-1a and SNAP-25. The SNAP-25 densitometric values in total extracellular vesicles fraction have been normalized to that of CD9 (a), while the STX-1a and SNAP-25 densitometric values in NDE have been normalized to that of NSE (c). (b, d, e) Densitometric analysis of STX-1a and SNAP-25 in extracellular vesicles The normalized SNAP-25 levels in T extracellular vesicles (b) and those of both STX-1a (d) and SNAP-25 (e) in NDE have been analyzed by using One-way ANOVA with post-hoc Tukey test. Both STX-1a and SNAP-25 expression levels are significantly increased in NDE of IS patients with respect to HC subjects (p ≤ 0.0001 and p ≤ 0.02 for STX-1a (d) and SNAP-25 (e) respectively), while no differences is observed for SNAP-25 in the T fraction (p = 0.91) (b). (g, h) Pearson’s correlation analysis between expression of STX-1a in IS NDEs and the clinical stroke. STX-1a levels in NDE of IS patients present a moderate positive correlation with NIH-SS (p = 0.003) (g) and a moderate negative correlation with BI (p = 0.02) (h). For WB analysis 50 µg of each extracellular vesicles fraction and 0.5 µg of mouse brain cortex lysate have been loaded in each lane. In quantitative graphs (panels b, d and e) each point in a frame depicts the value for a single subject (n = 10 IS patients and 10 HC subjects) while bars represent the median value ± SD. *p ≤ 0.05; ***p ≤ 0.001. In panels (b), (d) and (e) blue points and histograms correspond to HC subjects, red points and histograms correspond to IS patients. Uncropped WB in (a), (c) and (f) have been reported in Fig. S9 online.
