Figure 7.

Correlation of miR-451a levels in serum EVs with THP-1 macrophage response to inactivated WV. A, miR-451a mimic was transfected into THP-1 macrophages and was incubated for 2 days. EVs were collected from cell culture medium and then added to THP-1 macrophages. Cells were stimulated with inactivated WV for 6 h, and the expression of IFN-β mRNA was determined by RT-qPCR. Data represent means ± S.D. (n = 3) (p < 0.05, t test). B and C, THP-1 macrophages were cultured with each human serum (#1, #2, and #3), which was used in Fig. 5, for 2 days. Cells were stimulated with inactivated WV for 6 h, and the expression of IFN-β (B) and IL-6 (C) mRNA was determined by RT-qPCR. Data represent means ± S.D. (n = 3) (p < 0.05, t test). D, EVs were collected from 0.1 ml of human sera (#1, #2, and #3). Cells were cultured in serum-free medium with collected EVs in 24-well plates for 2 days and were subsequently stimulated with inactivated WV for 6 h. The expression of IFN-β mRNA was determined by RT-qPCR. Data represent means ± S.D. (n = 3) (p < 0.05, t test). E, antagomiR-451a was transfected into THP-1 macrophages, and cells were then cultured with or without human serum. IFN-β mRNA expressions in response to inactivated WV were determined by RT-qPCR. Data represent means ± S.D. (n = 3). “NS” represents “not significant” (p > 0.05, t test). F and G, schematic of treatment and preparation schedule (F). THP-1 macrophages were cultured with 20% of each human serum for 2 days. Cells were then stimulated with inactivated WV for 6 h, and IFN-β expression levels were determined by RT-qPCR. The expression levels of IFN-β mRNA were normalized to those of GAPDH. Correlation was investigated between miR-451a levels in serum EVs and type I IFN expression. Pearson correlation coefficient (r) was calculated, and a statistical analysis was performed (p < 0.05, n = 24, statistical power = 0.869) (G). The data are a representative of two independent experiments. *, p < 0.05.