Fig. 4

miR-484 directly targets SMAD2/ZEB1 and down-regulates their expressions. a The venn diagram shows the potential target genes (overlapping fraction) that were shared by all three databases. b The predicted binding sites for miR-484 in the 3′UTR of SMAD2 and ZEB1 and the mutations in the binding sites are shown. c The EGFP reporter assay was performed in HeLa or C33A cells co-transfected with pcDNA3/EGFP-SMAD2/ZEB1 3′UTR wild type or pcDNA3/EGFP-SMAD2/ZEB1 3′UTRmut with pri-miR-484 or ASO-miR-484. The fluorescence intensities were measured at 48 h after transfection. d Western blot assays were used to detect the SMAD2 and ZEB1 protein levels in HeLa or C33A cells transfected with pcDNA3/pri-miR-484, ASO-miR-484 or their corresponding controls. e Western blot assays tested the effect of SMAD2 on the expression of ZEB1. *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significance. All the bars indicate the means ± SDs. All experiments were performed at least triplicates