Verification of predicted schistosome miRNA targets in mammalian cells.

<p>Transfection of miR-2 (A), Let-7a (B), bantam (C), miR-8 (D and E), miR-31 (F), miR-1989 (G), and miR-3479 mimics (H) into HEK293T or Hela cells led to a significant reduction of luciferase activity from co-transfected plasmids containing their corresponding target regions. Hela or 293T cells were transfected with a recombinant Gaussia luciferase plasmid (pGLU-CMV) containing the corresponding <i>S</i>. <i>japonicum</i> cDNA fragment of a predicted mRNA targeted by a miRNA. These cells were simultaneously co-transfected with a control firefly luciferase plasmid. At 24 h post-transfection, a control miRNA or miRNA mimics were transfected into these cells. At 24–48 h post-transfection, luciferase activity was analyzed using a dual-luciferase reporter assay system with normalization to either firefly luciferase levels or protein concentration. For A and B, all the samples within the assay are illustrated, whereas in C-H only the effects of the miRNA mimics and scrambled miRNA mimics are shown. Similar results as illustrated in A and B were observed for the experiments in C-H. Blank represents no miRNA applied. Each experiment shows representative results and illustrates the mean and standard errors derived from triplicate experiments. * means P ≤ 0.05 and ** means P ≤ 0.01 (student’s t test, miRNA mimics treatment vs scrambled miRNA mimics treatment).