Fig. 3

Temporal clustering analysis of brain EV proteomes in AD. Fuzzy c-mean clustering of brain EV proteome was performed based on total number of spectra identified for each protein by label-free quantitative proteomics. Age-matched controls (control) were used as baseline in clustering analyses. Negative expression changes refer to lower concentration in AD compared to baseline. Proteins with low deviation were indicated by warm colored traces (red/magenta), while proteins with high deviation were colored in cold colors (blue/green/cyan). A total of 10 protein clusters were obtained in these analyses. a, c, e Cluster 8, cluster 7, and cluster 9, respectively (remaining clusters are displayed in Supplementary Figure 1 in the online-only Supplementary Material). Clustered proteins were further analyzed by one-way ANOVA with Bonferroni correction for multiple comparisons, unless otherwise specified. Significant differences are expressed versus controls. b, d, f Quantification of proteins significantly modulated from clusters 8, 7, and 9, respectively. *Significant differences P < 0.05. **Significant differences P < 0.01. ⁺Significant differences determined by ANOVA with Fisher’s LSD P < 0.05. ^#Significant differences determined by nonparametric one-way ANOVA on ranks P < 0.05. ^Significant differences determined by nonparametric one-way ANOVA with Holm-Šídák correction P < 0.05