Fig. 3

Characterization of EV isolated from patient plasma samples. A SEC isolated EV were examined for expression of common EV markers CD9, CD63, and CD81 via flow cytometry. Representative dot plots from a healthy control (HC) and RRMS patient. FSC and SSC was used to select events of appropriate size using the size gate determined as previously described. Events from the size gate were analyzed for expression for expression CD9, CD63, or CD81 via staining with corresponding anti-CD antibodies (anti-CD). EV stained with appropriate isotype control (IgG CT) were used as negative controls. Positive gates were determined using IgG-stained controls for each individual donor. B Representative results from a single donor for the percentage of size events positive for indicated surface markers when stained with anti-CD markers or isotype controls (IgG CT). Results represents the mean ± STDEV of four replicates. *** p ≤ 0.001 determined by t-test. C Percentage of size events positive for indicated surface markers from HC or RRMS patients. Results represents the mean ± STDEV of three separate donors. Each dot is an individual donor. ns = not significant determined by t-test. D Representative dot plots comparing size events positive for CD9 or CD63 isolated from a HC or RRMS patient. E Percentage of size events negative for CD9 and CD63 (CD9−, CD63−), positive for CD9 and negative for CD63 (CD9+ , CD63−), positive for CD9 and CD63 (CD9+ , CD63+) or negative for CD9 and positive for CD63 (CD9−, CD63+) from HC or RRMS patients. Results represents the mean of three separate donors. Percent of events were not significantly different between HC and RRMS patients as determined by t-test