Fig 2.

In vivo interaction between host miR-7 and viral wsv477 gene in shrimp. (a) Expression of miR-7 in shrimp. Either miR-7 or the LNA-modified anti-miR-7 oligonucleotide (AMO-miR-7) was injected into WSSV-infected shrimp. At different times (0, 12, 24, and 48 h) after injection, the shrimp gills were collected and subjected to Northern blotting with the miR-7-specific probe (upper panel). Shrimp U6 was used as a control. The bar chart in the lower panel represents the densitometric measurement of miR-7 expression in the Northern blot. The statistically significant differences between treatments are indicated with asterisks (*, P < 0.05; **, P < 0.01). (b) Effects of miR-7 on the expression of the viral wsv477 gene in Northern blotting. To detect the expression of wsv477, the shrimp specimens collected at 12 h after the injection of WSSV plus miR-7 or WSSV plus AMO-miR-7 were characterized by Northern blotting using the wsv477-specific probe. PBS and WSSV alone were used as controls. To evaluate the effects of miR-7 on the expressions of other viral genes, the mRNAs of wsv076 (an early gene of WSSV) and vp28 (a late gene of WSSV) were probed with gene-specific probes. The shrimp β-actin was used as a control. Lane headings indicate the solutions that were injected. The probes that were used are indicated on the right. (c) Effects of miR-7 on wsv477 expression in Western blotting. Western blot analyses were conducted with AP-conjugated goat anti-mouse IgGs using shrimp gills collected at 12 h after the injection of either PBS, WSSV, WSSV plus miR-7, or WSSV plus AMO-miR-7. The shrimp β-actin was used as a control. Lane headings indicate the solutions that were injected. The antibodies that were used are shown on the right.