Fig. 4

MiR-219-5p downregulated CCNA2 by directly targeting its 3’UTR. a Prediction of the target genes of miR-219-5p using online miRNA target prediction databases. Based on the list of the putative miR-219-5p binding sequences in the 3’-UTR of CCNA2, mutations were generated in the CCNA2 3’-UTR sequence in complementary sites for seed regions in miR-219-5p. b A human CCNA2 3’-UTR fragment containing either the wild-type or mutant miR-219-5p binding sequence was cloned downstream of the luciferase reporter gene. The luciferase activity was analyzed in 293 T cells. EC9706 and TE-9 cells were transfected with miR-219-5p or miR-NC and incubated for 48 h. The mRNA and protein levels of HMGA2 were measured using (c) real-time PCR and (d) western blotting. The data are presented as means ± SD of 3 independent experiments. **p < 0.01, ***p < 0.001, compared with miR-NC