Fig. 2.

miR-28 targets the 3′UTR of Nrf2 mRNA. a Schematics of Nrf2 mRNA 3′UTR and its potential miR-28 binding site. Nrf2 3′UTR mutant was generated with point mutations in the miR-28 binding site referred to as the MRE. b HEK293T cells transfected with wild type (WT) or mutant Nrf2 mRNA 3′UTR reporter plasmids with vehicle control or pre-miR-28. The luciferase activities were measured 48 h after transfection. The luciferase activities were normalized to those of renilla luciferase activity of a co-transfected reporter. The relative luciferase activities were calculated by normalizing to that of vehicle controls. c qRT-PCR showing the fold changes in mRNA levels after pre-miR-28 transfection of MCF-7 cells compared with control transfected cells. N = 2 ± SE, * P < 0.001. The expression level of normalized control cells is arbitrarily set as 1. d Western blotting showing Nrf2 protein expression in the pre-miR-28 transfected and vehicle-control transfected MCF-7 cells. β-actin was used as a loading control. A representative experiment was shown