Fig. 6

The effects of short-term activated HSC-sEV-specific miR-99a-5p on liver macrophage (KC) differentiation, inflammation and collagen deposition in a chronic liver injury mouse model. (A) Schematic representation of agomir-99a-5p treatment in a CCL4-induced chronic liver injury mouse model. (B) The expression of miR-99a-5p in KCs was determined by RT-qPCR and normalized to that of U6snRNA and to the miRNA agomir negative control (agomir-NC). (C) Surface marker expression in KCs gated on CD11b + F4/80 + primary liver mononuclear cells were determined by flow cytometry. Representative flow cytometry images and statistical histograms for CD86- and CD206-positive cells are provided. (D) Immunohistochemical staining of CD206 in liver tissue sections from each group. Representative images are provided, and positively stained cells are dark brown, scale bar = 100 μm. (E) The expression of NF-κB p65, IL-6, INOS, CD206, COLI, and α-SMA in the liver was detected by western blotting. β-Actin served as a loading control and was normalized to the mimic-NC treated group. Data from at least five mice for each group are presented as the mean ± SEM. Statistical significance was determined by one-way ANOVA, ***p < 0.001, **p < 0.01, *p < 0.05, ns, not significant