Figure 5

SATB2 was a direct target of miR- 182 in CRC cells. (A) Schematic illustration of the SATB2 3’UTR- containing reporter constructs. The mutant binding site was underlined. wt denotes wild type; mt denotes mutant. (B) Luciferase reporter assays in SW480 cells with cotransfection of wt or mt 3’UTR and miRNA as indicated. (C, D) Levels of SATB2 mRNA and protein after miR-182 ectopic over-expression in CRC cells were examined by real-time RT-PCR (C) and western blot (D). The immunosignal was quantified using Quantity One Software and relative protein abundance was determined by normalizing with β-actin. (E) A statistically significant inverse correlation was reported between miR-182 and SATB2 mRNA levels in CRC tissues (Spearman's correlation analysis, r = -0.7007; p = 0.001, n = 18).