Fig. 3

EZH2 is a direct target of miR-33a. a The bioinformatic methods used to predict the possible targets of miR-33a. b The predicted binding-sites of miR-33a in the 3′-UTR of EZH2 and its mutated version are shown. c, d Luciferase reporter assay was performed in MDA-MB-231 (c) and BT-549 (d) cells co-transfected with the pGL3 construct containing WT or MUT EZH2 3′-UTR region and miR-33a mimics. Data were normalized to those from cells co-transfected with the pGL3 and NC mimics. MDA-MB-231 and BT-549 cells were transfected with miR-33a mimics. e The EZH2 mRNA expression was detected and normalized to ACTB. f, g Quantitative analysis of Western blots is shown, and GAPDH served as a loading control. Data are shown as mean ± SEM (n = 3). ***P < 0.001 compared with cells transfected with NC mimics groups