Figure 2
H19 was a bona fide target for miR-141 and miR-22.
(A) Schematic diagrams of the mutual interplays between miRNA and H19. The positions of miRNA binding sites and calculated ΔG values are showed on the top (kcal/mol). (B) qRT-PCR results showed the relative RNA level of H19 after ectopic expression of miR-141 and miR-22. Neither miR-141 nor miR-22 affected the RNA level of H19. (C) HEK293 cells were transfected with miRNA mimics combined with luciferase reporter harboring H19 gene. The effects of miR-141 and miR-22 on the luciferase activity were measured by luciferase reporter assays. MiR-141 and miR-22 could suppress the luciferase activity. (D) The miR-141 and miR-22 binding sites were mutated and the mutated luciferase reporters were co-transfected with corresponding miRNAs. The mutations on binding sites abolished the previously suppressive effect. (E) H19 RNA level in the immunoprecipitates were measured by qRT-PCR. H19 RNA was enriched in the Ago2-coated beads. (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001).
