Fig. 4
hUC-EVs-ATO upregulated M2 macrophages in aGVHD mice. A On day 14 after transplantation, peritoneal macrophages were harvested from the mice and analyzed by flow cytometry. The frequencies of M1 (F4/80+CD86+) and M2 (F4/80+ CD206+) macrophages in the peritoneal fluid of mice that received PBS, ATO, hUC-EVs or hUC-EVs-ATO are presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, n.s. = not significant. There were six mice in each group. B On day 14 after transplantation, immunofluorescence images of M1 (F4/80+ iNOS+) and M2 (F4/80+ CD206+) macrophages in liver biopsy tissues and C intestinal biopsy tissues were obtained in mice that received PBS, ATO, hUC-EVs or hUC-EVs-ATO. Original magnification: × 400. Scale bar, 25 μm. D The number of M1 (F4/80+iNOS+) and M2 (F4/80+CD206+) macrophages in liver or intestinal biopsy tissues was quantified using a minimum of three mice in each group, with a minimum of six nonoverlapping fields of view per slide. Data represent the mean cell number per square millimeter ± SEM. E With CBA, the levels of TNF-α, IL-6, IL-1β, IL-10 and TGF-β in the serum of recipient mice in each group on day 14 after transplantation are described as the mean ± SEM. *P < 0.05; **P < 0.01, ***P < 0.001, n.s. = not significant. There were six mice in each group