Figure 3
Direct targeting MMP1 3′ UTR of miR-526b. (a) The nucleotide resolution of the predicted target sites (377–383 position) of the MMP1 3′ UTR and mature miR-526b sequence: the seed sequence (blue letters); the target sequence (red letters); and the evolutionarily conserved regions (gray boxes). All of the sequences for mature miR-526b from human (hsa), chimpanzee (ptr), rhesus monkeys (mml) and orangutan (ppy) were perfectly conserved. The predicted target sequences of the MMP1 3′ UTR were only conserved in primates, including humans, chimpanzees and rhesus monkeys. (b) A sketch of the construction and sequence alignment of the wild-type and mutant MMP1 3′ UTR with hsa-miR-526b is shown. The mutated sequences are underlined. (c) The wild-type or mutant MMP1 3′ UTR reporter vector was transfected into JAR cells. The luciferase and β-galactosidase activities were measured after 72 h. The luciferase activities were normalized to the β-galactosidase activities. (d) The wild-type or mutant MMP1 3′ UTR reporter vector was co-transfected into the HeLa cells with either the N.C. mimic or miR-526b mimic. The luciferase and β-galactosidase activities were measured after 72 h. The luciferase activities were normalized to the β-galactosidase activities. The data are shown as the mean±s.e.m. of at least three independent experiments (*P<0.05).
