<p>(A) Nucleotide sequences and modifications of the anti-miRNA-1 oligonucleotide (AMO-miR-1) and AMO-miR-1-scrambled. (B) Silencing of miR-1 expression in shrimp hemocytes <i>in vivo</i>. The miR-1-specific AMO (AMO-miR-1) and the negative control AMO-miR-1-scrambled were injected into shrimp, respectively. The shrimp hemocytes were subjected to Northern blot using miR-1 or U6 probe. (C) The effect of miR-1 expression silencing on phagocytic activity of shrimp hemocytes. The phagocytic activity of FITC-labeled WSSV was evaluated with flow cytometry. The plotted data points referred to the means ± standard deviations of triplicate assays and the asterisk represented statistically significant differences (*, <i>p</i><0.05). (D) The interaction between miR-1 and <i>CLTC1</i> gene. The miR-1 precursor and the plasmid EGFP-<i>CLTC1</i> or EGFP-<i>CLTC1</i>-mutant or EGFP were cotransfected into insect High Five cells. Then the fluorescence intensity of cells was monitored with a fluorescence microscope. The columns represented the means ± standard deviations of triplicate assays. The significant differences between treatments were indicated with asterisks (**, <i>p</i><0.01). (E) The effect of <i>CLTC1</i> on phagocytosis of shrimp hemocytes. The shrimp were injected with <i>CLTC1</i>-siRNA or <i>CLTC1</i>-siRNA- scrambled as a control. Then the shrimp were subjected to Northern blots (left) and phagocytosis assays (right). In Northern blots, the shrimp β-actin was used as a control. The significant differences between treatments were indicated with asterisk (*, <i>p</i><0.05). (F) Sequence alignment of miR-1 from six typical species. * indicated the identical nucleotides.</p
