Fig. 6

ROR-EVs are novel biomarkers for breast cancer. A Representative NTA of lEVs and sEVs isolated from peripheral blood of a breast cancer (BC) patient. B lEVs from four BC patients were characterized by western blot for the expression of common lEV markers (α-actinin-4, RGAP1, ꞵ-Actin) or lipoprotein contaminants (ApoA1, ApoB). C Flow cytometry of patient-derived lEVs including PBS + 1%EV-depleted FCS, ROR2 antibody-only or size beads as controls. D + E The percentage of lEVs carrying the tumor-related antigens ROR1, ROR2, or EpCAM isolated from healthy controls (CTL) or BC patients was measured by flow cytometry. Shown are representatives histograms from one breast cancer patient for each marker (D) as well as the quantification from all analyzed samples (E). Boxes mark the 25–75 percentiles (line at median) and whiskers the 5–95 percentile. Significance was calculated with a Mann–Whitney test. F + G ROC analyses of EV-associated ROR1, ROR2 and EpCAM alone (E) or of all three markers combined (F)