BMDC-EV attract immune cells in lymph nodes similarly as in the skin. BMDC-EV attract immune cells in lymph nodes similarly as in the skin. (A) Individual images of a MELC analysis, assessing EV and immune cell markers in lymph nodes. Tissue sections from draining lymph nodes after skin injections of imBMD-EV and maBMD-EV (red label) obtained after 24 h were analyzed by MELC for the indicated markers and for colocalization with EV. Numbers in the left-most panels depict the localization of cells analyzed in (D). (B) The relative presence of cells in EV-containing areas (in percentage) was assessed by StrataQuest software as in Figs 4 and 5. The analysis was performed in tissue sections from four different lymph nodes. The respective numbers served to calculate the SEM. (C) Monocytes in imDC-EV and maDC-EV areas. Monocytes (CD11b+/Ly6C+/Ly6G−) were quantified in the EV areas using the StrataQuest software as explained in Table 1 and Fig S8A. (D) Identified monocytes were analyzed for co-expression of additional markers, expressed in percentage of total monocytes found in the EV area. The analysis was performed in tissue sections from four different injection sites. The obtained numbers were used to determine the SEM. Scale bars represent 100 μm. (E) Expression levels of indicated markers for each cell numbered in (A). For better understanding, protein expression was divided into three levels (no, low, and high expression), which were color-coded as in Fig 6B. Scale bars represent 100 μm. Stefan Schierer et al. LSA 2018;1:e201800093 © 2018 Schierer et al.