FIGURE 4:

Changes in activities of Smurf2 3′-UTR luciferase reporters after deletion of miR-322/503–binding site. (A) Effect of 5′ deletion of Smurf2 3′-UTR on its luciferase reporter activity in cells transfected with pre–miR-322 (a) or pre–miR-503 (b). Top, schematic of plasmids of different chimeric firefly luciferase Smurf2 3′-UTR reporters. BS, predicted miR-322/503–binding site. Bottom, Smurf2 3′-UTR luciferase reporter activity. At 24 h after transfection with pre–miR-322 or pre–miR-503, cells were cotransfected with Smurf2 F-Luc constructs and a Renilla luciferase reporter. Firefly and Renilla luciferase activities were assayed 24 h later. Results were normalized to the Renilla luciferase activity and expressed as means ± SEM of data from three separate experiments. *p < 0.05 compared with cells transfected with control scrambled oligomer. (B) Effect of point mutation of specific miR-322/503–binding site (schematic) in Smurf2 3′-UTR on luciferase reporter activities after ectopic overexpression of miR-322 (left) or miR-503 (right). *p < 0.05 compared with cells transfected with control scrambled oligomer.