Fig. 3

miR-195-5p affects colon cancer cell EMT by modulating NOTCH2 expression in a post-transcriptional manner. Western blots of NOTCH2, Ad-NOTVH2, and EMT markers Vimentin and E-Cadherin in DLD1 and HCT116 cells transfected with different miRNAs (b) and quantifications of NOTCH2 protein (c). Assays were performed in triplicates. A schematic representation of the pmiR-RB-REPORTTM dual-luciferase reporter vector with 3′-UTR of Notch2 mRNA harbors two miR-195-5p cognate sites (d). Relative luciferase activity of reporter plasmids carrying wild-type or mutant Notch2 3′-UTR in DLD1 and HCT116 cells co-transfected with miR-195-5p mimic or mimic negative control (mimic NC) (e). Notch2 mRNA decay curves of HCT116 cells carrying miR-195-5p mimic or mimic NC (f). The decay model was fit in a one-phase exponential decay model. AGO2-RNA immunoprecipitation assay (RIP) showing that miR-195-5p interacted with NOTCH2 in HCT116 cells (g). (**P < 0.01, ***P < 0.001)