Fig. 6

Multiplex antibody bead-based surface profiling analysis of EVs reveals an immuno-modulatory signature. a Representative flow cytometric dot plots of 37 surface epitopes plus two isotype controls (39-bead plex) are shown for each of the four hBM-MSC donors (81RB/48RB/55RB/85RB; panels i–iv, respectively). The flow cytometric plots represent the 39-bead gating areas representative of each epitope analyzed with its own gate according to the calibration bead distribution (top panels). Positive EV populations were detected using a CD63-APC detection antibody (lower panels) and double positive (specific epitope: CD63-APC) populations are shown in the multiplex analysis. b Quantification of the flow cytometric results obtained by the MACSPlex multiplex analysis for each of the EV samples harvested at three time points of collection (days 1, 13, and 25) and of all four hBM-MSC donors (N = 4; hBM-MSC-EV-81RB/48RB/55RB/85RB). Data are showing the mean for each time point of collection and standard error of mean (S.E.M) represent the four hBM-MSCs. Statistical significance was analyzed by a two-way analysis of variance (ANOVA) with Tukey’s post hoc analysis. A p value of < 0.05 was considered statistically significant and significant differences are marked with a single (p < 0.05), double (p < 0.01), triple (p < 0.001), or quadruple (p < 0.0001) asterisks