Figure 4.

miR-135b Directly Targets TET3 and Multiple Negative Regulators of the Wnt/β-Catenin Pathway

(A) Predicted binding sites of miR-135b in the 3′ UTR of the indicated genes. (B and C) qRT-PCR (B) and western blot analysis (C) of the expression of the indicated genes in the cells shown. (D) RIP analysis revealed that mRNAs of the indicated genes were recruited to miRNA-protein (miRNAP) complexes following immunoprecipitation with flag (flag-Ago2). IgG immunoprecipitation was used as a negative control. (E) Activity of the luciferase gene was linked to the 3′ UTRs of the indicated genes. The luciferase reporter plasmids (wild or mutated 3′ UTR sequences of the indicated genes) were transfected into the indicated cells, and the luciferase activity was measured 48 hr after transfection. (F) The indicated cells were transfected with the indicated genes and a TOP or FOP reporter and were then subjected to a dual-luciferase assay 48 hr after transfection. (G) The indicated cells were transfected with the indicated genes and were subjected to western blot analysis 72 hr after transfection. *p < 0.05; **p < 0.01; ***p < 0.001. nd, not detected.