Figure 3.

miR-9* targets the 3′ UTR of CoREST. A, Cotransfection of HEK 293 cells with luciferase reporters containing the 3′ UTR of CoREST in the presence of increasing concentrations (0.1, 1.0, 5.0, or 15.0 nm) of artificial pre-miR-9* reduces relative luciferase activity (n = 5). B, C, CoREST levels are reduced following transfection of HEK 293 cells with pre-miR-9* (30 nm). A representative Western blot (B) and densitometry (n = 4) (C) after normalized β-actin levels. Data are mean ± SEM; *p < 0.05, **p < 0.01. D, TSP designed the predicted miR-9* binding site of CoREST (TSP +) rescues pre-miR-9* mediated knockdown (B). HEK 293 cells were cotransfected with pre-miR-9* (15 nm) and TSP+ or TSP Neg and CoREST levels measured in cell lysates by Western blot 24 h later. Densitometry from n = 4 independent experiments indicate increased CoREST in TSP+ (1.02 ± 0.05)-treated versus TSP Neg (0.565 ± 0.08)-treated cells.