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. 2013 Oct 8;8(1):83–92. doi: 10.1016/j.molonc.2013.09.004

Figure 5.

Figure 5

miR‐203 regulates ATM protein expression by directly binding to the ATM mRNA 3′UTR. A. Site‐directed mutagenesis targeting two potential miR‐203 binding sites (mutants 1 and 2) on an ATM mRNA 3′UTR‐luciferase construct. The ATM 3′UTR was studied in two forms (a and b) to cover the full length of the 3′UTR region. B. Luciferase activities of ATM mRNA 3′UTR‐luciferase constructs containing either the wild type or a deletion mutant for miR‐203 binding sites after co‐transfection of either pre‐miR‐203 or anti‐miR‐203 in 293 cells. miR‐203 significantly decreased the ATM 3′UTR (form b) luciferase activity, and miR‐203 inhibitor significantly enhanced the ATM 3′UTR (form b) luciferase activity. Both effects were blocked by mutant 1, but not by mutant 2. C. ATM protein expression levels are regulated directly by miR‐203, as reflected by decreased ATM expression in HT29 cells after transient transfection of pre‐miR‐203 and increased ATM expression in HT29‐OxR cells after miR‐203 inhibitor transfection. *P < 0.05. Results are representative of three experiments.