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. 2017 Nov 27;8:1546. doi: 10.3389/fimmu.2017.01546

Figure 4.

Figure 4

Simultaneous interaction between a microRNA (miRNA) and its two targets. (A) The miRNA-mediated degradation of target mRNA. White spot syndrome virus (WSSV)-miR-N32 at different concentrations was incubated with Ago1 complex and wsv459-3′UTR or wsv322-3′UTR for 2 h. Then the wsv459 mRNA or wsv322 mRNA was detected by Northern blot. (B) The simultaneous targeting of two mRNAs by WSSV-miR-N32. WSSV-miRN32 at various concentrations was incubated with the mixture of Ago1 complex, wsv459-3′UTR and wsv322-3′UTR for 2 h. Subsequently, the wsv459 and wsv322 mRNAs were detected by Northern blot. (C) The co-localization of WSSV-miR-32 and wsv459 mRNA in the hemocytes of WSSV-infected shrimp. The shrimp were infected with WSSV. At 6 and 24 h postinfection, shrimp hemocytes were labeled using WSSV-miR-32-specific and wsv459 mRNA-specific fluorescent probes. Scale bar, 10 µm. (D) The co-localization of WSSV-miR-32 and wsv322 mRNA in the hemocytes of WSSV-infected shrimp. Scale bar, 10 µm. (E) The co-localization of WSSV-miR-32, wsv459 mRNA, and wsv322 mRNA in the hemocytes of WSSV-infected shrimp. Scale bar, 10 µm.