Total Exosome protein isolated by this procedure was low and variable (18 – 131 (mu)g). This variability was described by Knepper et al., who noted significant numbers of exosomes were lost in the uromodulin complex. MudPIT analysis was done on 50 (mu)g protein, or the total isolated, whichever was greater. Confident protein IDs were substantially greater than that reported previously. In this work we identified 2,059 protein groups with more than 15 spectral counts in both groups. Aldosterone treatment caused a statistically significant change (P < 0.05) in 395 proteins (of 2,059). The top 100 most abundant proteins are reported in Table 2. RNA analysis by deep sequencing yielded 26,803,283 reads that were subsequently mapped to 12,833 transcripts (genes or miRNAs). Gene Ontology classification of exosomal protein and mRNA showed a similar pattern of expression in both groups. However, further work is required to evaluate the effects of aldosterone on gene expression. Candidate biomarkers will be selected for further study from a list of the top 100 most abundant proteins affected by ALDO treatment (P < 0.05, Student’s paired ttest).
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