Figure 4

Quantitative analysis of STX-1a in PBMCs and of SNAP-25 in sera in all population analyzed. (a, e) Representative WB analysis of STX-1a in PBMCs (a) and of SNAP-25 in serum (e) in IS and HC subjects. PBMCs are immunostained for STX-1a (a) and serum for SNAP-25 (e). For each sample, the STX-1a densitometric value is normalized to that of β-actin, while the SNAP-25 densitometric values is normalized to that of total blood proteins content revealed by Coomassie Blue staining. (b) Densitometric analysis STX-1a expression levels in PBMCs. STX-1a expression levels have been analyzed by using One-way ANOVA with post-hoc Tukey test: STX-1a significantly increase in PBMCs of IS patients with respect to HC and Y-HC subjects (p ≤ 0.0001), while no differences are observed between control groups (p = 0.97). (c, d) Pearson’s correlation analysis between expression of STX-1a in IS PBMC and the clinical stroke scales. STX-1a levels in PBMCs of IS patients present a moderate negative correlation with NIH-SS (p = 0.03) (c) and a moderate positive correlation with BI (d) (p = 0.03). (f) Densitometric analysis SNAP-25 expression levels in sera. SNAP-25 expression levels have been analyzed by using One-way ANOVA with post-hoc Tukey test: SNAP-25 levels are significantly increased in serum of IS patients with respect to HC and Y-HC subjects (p ≤ 0.0001). A significant increase of SNAP-25 expression is also observed in HC with respect to Y-HC subjects (p ≤ 0.0001). (g) WB analysis of SNAP-25 in sera depleted of IgG and albumin. The increased SNAP-25 levels in IS patient are not influenced by the presence of serum proteins. For WB analysis 100 µg of whole PBMCs and of serum (SE) have been loaded in each lane. In quantitative graphs (panels b and f) each point in a frame depicts the value for a single subject (n = 30 IS patients, 30 HC subjects and 15 young HC subjects) while bars represent the median value ± SD. ***p ≤ 0.001. In panels (b) and (f) light blue points and histograms correspond to Y-HC subjects, blue points and histograms correspond to HC subjects, red points and histograms correspond to IS patients. Uncropped WB and coomassie staining in (a), (e) and (g) have been reported in Fig. S7 online.