EZH2 as a potential miR-26a target. (A) Protein expression analysis of selected miR-26a candidate targets in stably transfected Raji cells at indicated time points after miR-26a induction compared with an empty vector control. Actin/tubulin served as a loading control. (B) Transient transfection of miR-26a precursor and scrambled RNA into HEK-293 and Raji cells. Western blot results are shown for EZH2 and tubulin 48 hours after transfection. (C) Protein expression analysis of EZH2 after Dox treatment in 2 MYC-induced murine lymphoma cell lines. Starvation of Namalwa cells served as a positive control. (D) EZH2 protein expression in several human BL cell lines compared with endogenous miR-26a expression. (E) Sequence homology between miR-26a and human EZH2, site of miR-26a binding in the 3′-UTR of EZH2 is indicated. (F) Fold repression of luciferase activity as determined by reporter assay in HEK-293 cells. Luciferase expression was determined 24 hours after cotransfection of either the precursor miR-26a or a scrambled RNA. There was a significant reduction in luciferase activity in the miR-26a transfected cells compared with the controls (P = .004, exact Wilcoxon rank sum test). Error bars for 3 independent experiments (measured each in triplicate) are depicted.