Fig. 7

Short-term activated HSC-sEV-specific miR-99a-5p-educated macrophages suppressed the growth of hepatocellular carcinoma. THP-1 macrophages were pretreated with miR-99a-5p mimic or mimic negative control (mimic-NC) for 8 h. (A) Colony formation of Huh7 or HepG2 cells cocultured with miR-99a-5p mimic-pretreated THP-1 macrophages at a ratio of 5:1. (B) Nude mouse xenograft experiments. HepG2 cells were mixed and subcutaneously cotransplanted with miR-99a-5p mimic-pretreated THP-1 macrophages. The size of the tumor was measured at the indicated time points. The weight of the tumor was obtained on Day 28. (C) Immunohistochemical staining of Ki67 in paraffin sections of xenografts, scale bar = 100 μm. (D) The expression of CD206 and INOS in xenografts was detected by western blotting, β-Actin served as a loading control. For (A-D), the experiments performed with THP-1 macrophages pretreated with miRNA, mimic-NC served as a negative control. Representative images are provided. Data from at least three wells (A) or five mice (B-D) for each group were shown as the mean ± SEM. Statistical significance was determined by one-way ANOVA, ***p < 0.001, **p < 0.01, *p < 0.05, ns, not significant