Figure 3

Expression of MET was regulated by miR-144. (A) mRNA levels of putative miR-144 targets were examined by real-time PCR in SNU-5 cells transfected with miR-144 mimics or miR-NC. (B) Protein levels of putative miR-144 targets were examined by western blotting in SNU-5 cells transfected with miR-144 mimics or miR-NC. (C) The % input recoveries of the RNA-ChIP reactions illustrates the enrichment by MET antibody. RNA-ChIP assay for miR-144 performed on anti-MET antibody from lysates of cells. RNA-ChIP with a nonrelated IgG served as controls. (D) A schematic picture of the predicted miR-144 binding site in the 3′UTR of MET. (E) SNU-5 cells were transiently co-transfected with LUC-MET 3′UTR and miR-144 mimic. (F) AGS cells were transiently co-transfected with LUC-MET 3′UTR and a miR-144 inhibitor. (G) Mutating of the miR-144 binding site in MET 3′UTR abolished the miR-144-induced luciferase activity suppression. Luciferase activity was measured after 24 h and normalised to the co-transfected Renilla. (*p < 0.05; **p < 0.01; ***p < 0.001).