The MDM4 open reading frame contains a functional miR-34a responsive site.

<p>(A) Binding site for miR-34a predicted in human MDM4 mRNA by miRanda. Binding is indicated by solid lines, while wobble base pairing is indicated by a dashed line. Identity with the human MDM4 sequence is indicated by asterisks. The seed region of miR-34a, positions 2–7, are boxed. The homologous region of human MDM2 mRNA is also shown for comparison. (B) In MCF7 cells, the miR-34a inhibitor was cotransfected with a reporter gene containing the ORF miR-34a site from MDM4 (“Exon 11″) or the reporter with the rs79824231 SNP (“Exon 11 A>C”). Data are the averages of at least six independent experiments, with standard deviation indicated in error bars. Asterisks indicate t-test values <0.5. (C) In H1299 cells, the reporters were cotransfected with an expression plasmid for miR-34a or miR-34a-mut. Data are the averages of at least six independent experiments, with standard deviation indicated in error bars. Double asterisks indicates paired, one-tailed t-test value <0.01.