Fig. 4

MMP2 serve as functional target gene of miR-199a-5p. A The intersection of miRWalk, ENCORI, miRPathDB and miRDB to predict miR-199a-5p targets. B-C Western blotting and qRT-PCR validated of the predicted results that miR-199a-5p significantly restrained the expression of MMP2 at transcription and protein expression level. D The binding sites of MMP2 were predicted by the ENCORI database and the plasmids for the dual luciferase assay were designed. E The dual luciferase reporter assays revealed that the luciferase activity decreased after co-transfecting Luc-MMP2-wt with miR-199a-5p mimics. F The Image utilizing the confocal microscopy showed the binding of miR-199a-5p to MMP2 mRNA (the yellow points) in the cytoplasm. G The AGO2 recognition of miRNA binding complex containing MMP2 was measured by AGO2-RIP. H-I MMP2 mRNA was enriched in the AGO2-beads. MMP2: matrix metalloproteinase-2; NC: negative control; DAPI: 4’,6-diamidino-2-phenylindole; qRT-PCR: quantitative real-time polymerase chain reaction. ns: P > 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P < 0.0001