Figure 2

miR-143 directly targets MACC1 by binding to its 3'UTR. (A) The predicted miR-143 binding site within MACC1 3'UTR and its mutated version by site mutagenesis are as shown. (B) Variable MACC1 expression in six CRC cell lines was obtained by Western blot. β-actin was used as the loading control. (C) Non-metastatic SW480 cells expressed significantly higher level of miR-143 compared with its metastatic counterpart SW620 cells (* P < 0.05). (D) The repression of luciferase activity by MACC1 3'UTR was dependent on miR-143. Mutated MACC1 3'UTR abrogated miR-143 mediated repression luciferase activity (** P < 0.01). (E) In comparison with scramble controls, elevated expression of miR-143 by mimics inhibited MACC1 expression at both mRNA and protein level, while reduction of miR-143 by inhibitors moderately restored MACC1 expression (* P < 0.05). Figure is representative of 3 experiments with similar results.