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. 2021 Aug 27;17(14):3745–3759. doi: 10.7150/ijbs.57610

Figure 5.

Figure 5

Inhibition of miR-9-5p shuttled by NAFLD-EVs relieves macrophage inflammation. Macrophages were treated with EVs from the plasma of healthy individuals or NAFLD-EVs or control-EVs and PA-EVs, and then transduced with In-NC and miR-9-5p Inhibitor. THP-1 cells were treated with 100 ng/mL PMA for 24 h to facilitate their differentiation into macrophages. PKH67-labeled EVs were incubated with THP-1 cells at 37℃ for 12 h and the following indicators were then determined: A, B, miR-9-5p level in macrophages measured by RT-qPCR. C, D, TGM2 protein level in macrophages measured by Western blot analysis. E, Levels of IL-1β, IL-6, TNF-α and iNOS in macrophages measured by RT-qPCR. F, Positive rate of CD86+ CD11b+ elevated in macrophages detected by flow cytometry. G, Expression of inflammatory factors in supernatant of macrophages determined by ELISA. N = 3. * p < 0.05 vs. macrophages transduced with Control-EVs/No NAFLD-EVs; # p < 0.05 vs. macrophages transduced with PA-EVs + in-NC/NAFLD-EVs + in-NC. Data are shown as the mean ± standard deviation of three independent experiments. Data among multiple groups were compared using one-way ANOVA with Tukey post hoc tests used.