Identification of Itch as a target of miR106b in primary leukemia cells. (A) Primary leukemia cells were transfected with 300 nM precursor molecules for miR106b, nontargeting pre-miRNA control (scrambled) or left untreated for 72 hours. The expression levels of Itch, p73, and actin were evaluated. (B) Levels of miR106b quantitated by RT-PCR in leukemia cell samples transfected with miR106b or scrambled sequences as described in panel A. (C) Expression of Itch mRNA as normalized to GAPDH (C) and Itch protein as normalized to actin (D). Error bars represent SD from transfections of 2 independent leukemia samples analyzed in duplicate. One-way analysis of variance was performed for the quantified values of Itch protein under different transfection conditions with the Prism 5 software. ** Statistical significance (P < .05).