Tables and Figures
Figure 1: Rivaroxaban modulates a tightly regulated cluster of proteins collectively involved in the regulation of coagulation.Platelet poor plasma samples from individual Rivaroxaban-treated (n=6) and warfarin-treated patients (n=6) were enriched for vesicular fractions by ultracentrifugation. (A) 20 µg protein was separated on a 10% SDS-polyacrylamide gel. Immunoblotting for transmembrane (CD63 (1:1000) and CD81 (1:1000)) and soluble (HSP70 (1:1000)) EV markers confirmed the presence of vesicles in the enrichments across two randomly chosen donors in each cohort. Detection of albumin (1:1000) indicated co-isolation of plasma proteins. (B) Proteomic analysis in technical duplicate revealed a total of 181 proteins identified in at least 50% of at least treatment group, with one protein (PROZ - Vitamin K-dependent protein Z) exclusive in the Rivaroxaban cohort. (C) Volcano plot comparing the expression level of the 180 shared proteins between the Rivaroxaban versus warfarin vesicular proteomes (x-axis, Welch’s test difference representing the difference between the mean log2 LFQ values of Rivaroxaban to warfarin EV proteomes; y-axis, the negative log transformed p-value), representing the proteins significantly altered between the two cohorts. The black hyperbolic line indicated the threshold for statistical significance using an FDR of 0.05 and S0 of 0.1. Of the 180 proteins shared between the cohorts the expression of 5 proteins was significantly increased in the Rivaroxaban cohort (red). The expression level of the remaining 175 proteins remained unaltered (grey). (D) STRING functional protein association network analysis (version 11.5) of the differentially expressed and unique proteins revealed a tight protein-protein interaction network attributed to blood coagulation (red,p =1.43x10-7).