1. EV derived from mature BMDC are efficiently taken up by murine monocytes and have a rich content of CCF.
EV derived from mature BMDC are efficiently taken up by murine monocytes and have a rich content of CCF. (A) Murine PBMCs were incubated with stained (PKH26) immature (im) or mature (ma) BMDC-EV for 2 h. Percent of PKH26+ cells was determined by flow cytometry gating on monocytes (Ly6C-high, CD11b+, and Ly6G−), B cells (B220+, CD11b−, Ly6C−, and Ly6G−), and granulocytes (SSC-high, Ly6G+, CD11b+, Ly6C-int., and B220−). (B) EV derived from matured BMDC contain a rich CCF. EV derived from mature BMDC, which had been stimulated with Poly IC/R848 (EV-BMDC + Poly I:C) or LPS (BMDC-EV + LPS) or unstimulated (BMDC-EV), were analyzed by an antibody-based bead array (BioLegend). Values were normalized to number of EV-producing cells. Five different donors were analyzed. The resulting numbers were used to calculate mean values ± SEM. (C) Lysates from EV as described in (B), but derived from cells of a different donor animal, were analyzed by a protein array as described in Fig 3. The pixel intensity of each dot was determined by ImageJ, and the value was adjusted in relation to the internal positive control, which was set to 1. Shown is one representative analysis performed with four different donors.
Stefan Schierer et al. LSA 2018;1:e
© 2018 Schierer et al.