Figure 1

miR-29 targets NMI . (A) Micro-RNAs predicted to target the NMI transcript are catalogued using three different data bases represented as distinct columns. The top hits that were common to at least two data bases are represented. The highlighted miRNAs are common to all 3 searches. The miR-29 family is represented by underline. (B) Alignment of the miR-29 a, b, c seed sequence with the NMI 3′ UTR. Dashed lines represent complementary base pairing. (C) pMIR-REPORT29-NMI (containing putative binding site of miR-29 from the 3′ UTR of NMI) was co-transfected with pre-miR-29 or scrambled control. The assay was performed in triplicate and the experiment was performed twice. The luciferase activity readings were normalized with activity from a co-transfected β-gal expressing control. The error bars represent standard error of the mean (SEM). [* indicates p ≤0.05]. (D) Levels of each of the miR-29a and b mature transcripts from human breast cancer cell lines compared to MCF7, a breast cancer cell line with distinct epithelial like characters using real time-quantitive-PCR. Reactions were performed in triplicate. The corresponding level of NMI protein is seen in the adjacent western blot. GAPDH levels were used as loading control. (E) Transient transfection of MCF7 cells with pre-miR-29 a or b reduced NMI protein levels compared to scrambled control. On the other hand, MDA-MB-231 cells showed elevated levels of NMI upon introduction of anti-miR-29 a or b.