Figure 6

The BCL11A 3′-UTR is the target of miR-30a, but not of miR-1. pmirGLO-BCL11A luciferase constructs, containing a wild-type or mutated BCL11A 3′-UTR (A), were transfected into A549 cells with (B) miR-30a, (C) miR-1, or control miRNA vectors. Relative repression of firefly luciferase expression was standardized to Renilla luciferase activity. The reporter assays were performed three times with essentially identical results. The P value was calculated comparing control microRNA and miR-30a or miR-1. (D) miR-30a downregulated BCL11A protein expression in H520 cells. Control miRNAs were also used for transfection. The experiment was performed three independent times, and representative results are displayed. CD511B represents the pCDH-CMV-MCS-EF1-copGFP empty vector. (E) Immunofluorescence staining intensity of BCL11A protein in H460 cells observed by confocal microscopy after transfection with miR-30a or control miRNA. Magnification, ×200.