Figure 4.

Circulating Exosomes from Mice with EAM Are Selectively Loaded with miR-142

(A) The expression of inflammatory miRNAs in serum exosomes from control and EAM groups was measured by qPCR. (B) CD4⁺ T cell miRNA levels were measured by qPCR after incubation with Con-Exos and EAM-Exo. (C) Correlation between the expression of miR-142 in exosomes and expression in splenic CD4⁺ T cells was analyzed. (D) Exosomes were added to CD4⁺ T cells in the presence or absence of 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole, an inhibitor of RNA polymerase II transcription. After 24 h, we analyzed the miR-142 level in recipient cells. (E–L) CD4⁺ T cells were transfected with the miR-142 inhibitor or inhibitor control (NC) for 24 h and then treated with EAM-Exo. (E) miR-142 levels were measured after transfection and exosome incubation. (F) CD25 and CD69 expression in respective groups was measured. (G) T cell proliferation was determined based on BrdU incorporation. (H) Percentages of Th1, Th17, and Treg cells were analyzed by flow cytometry. (I) Levels of the cytokines IFN-γ, IL-17A, and IL-10 were measured by ELISA. (J) ECAR in CD4⁺ T cells was calculated. (K and L) Cellular glucose uptake (K) and lactate production (L) were measured. Data are presented as the means ± SEM (n = 6); ∗p < 0.05.