Fig. 3

Exosomes mediated the EC-macrophage crosstalk. A Schematic diagram of the EC-macrophage co-culture system, upper layer: ECs; lower layer: macrophages. B Western blotting analysis of the expression of iNOS (M1 macrophage marker) and Arg-1 (M2 macrophage marker) in Vehicle, UTX^f/f ECs, and UTX^−/− ECs with the addition of GW4869. C Statistical analysis of iNOS and Arg-1 expression in each group in B. D–I qRT-PCR analysis of the mRNA expression of M1 markers (iNOS2, TNF-ɑ and CD86) and M2 markers (Arg-1, IL-10 and CD206) in Vehicle, UTX^f/f ECs and UTX^−/− ECs with addition of GW4869. J Representative scatter plot of WT mice taken up Dil-labeled exosomes on day 7 after SCI. Cells were immunolabelled with CD45-APC and CD11b-PerCP-Cy5.5 antibodies. Dil⁺ cells are defined as the cells taken up Dil-labelled ECs-Exos, macrophages defined as CD45^highCD11b⁺, and microglial cells defined as CD45^lowCD11b⁺. The numbers in the gates refer to the percentage of positive cells for each marker. K Statistical analysis of the percentage of macrophages, microglia, and other cells taken-up Dil-labelled ECs-Exos in CD45 and CD11b gate. L Representative scatter plot of macrophage uptake of Dil-labeled ECs-Exos on day 7 post-SCI. The numbers in the gates refer to the percentage of positive cells for each marker. M Statistical analysis of Dil⁺ macrophages and Dil⁺ microglia percentage in Dil⁺ gate. ^nsP > 0.05, *P < 0.05, **P < 0.01, compared with corresponding control mice. n = 5/per group