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).