Identification of Itch as a target of miR106b in K562 cells. (A) Complementary binding between miR106b and the 3′UTR of Itch. (B) Relative luciferase activity of luciferase reporter constructs containing full-length and either the antisense or mutated construct (values shown together as pGL-Itch-UTR mut) on addition of 300 nM of the precursor molecules for miR106b or scrambled miRNA. (C) Quantitation of miR106b expression in K562cells transfected with 300 nM precursor molecules for miR106b, miR130a (an unrelated miR not predicted to target Itch), anti-miR106bAM, anti-miR130aAM, and a nontargeting pre-miRNA control (scrambled) or left untreated for 48 hours. (D) The levels of Itch mRNA were quantitated after normalizing to GAPDH in the same samples. (E) Immunoblotting was performed for Itch and GAPDH in the same samples.