Figure 3.

MiRNA-324-5p activates Wnt/β-catenin signaling pathway and EMT. (a)MiRNA-324-5p activated Topflash luciferase activity. BGC-823 cells were co-transfected with NSC-inh or miRNA-324-5p inhibitors, Topflash/Fopflash plasmids and pTK-renilla. *p < 0.05. (b) MiRNA-324-5p activated Topflash luciferase activity. HFE-145 cells were co-transfected with NSC-mim or miRNA-324-5p mimics, Topflash/Fopflash plasmids and pTK-renilla. *p < 0.05. (c) Suppression of miRNA-324-5p inhibited the translocation of β-catenin into the nucleus. Forty-eight hours after miRNA-324-5p inhibitors transfection, BGC-823 cells were lysed and immunoblotted with β-catenin antibody in cytoplasmic and nuclear portion, respectively. (d) Inhibition of miRNA-324-5p blocked the translocation of β-catenin into the nucleus. β-catenin localization was detected by confocal immunofluorescence 48 h after transfection of miRNA-324-5p inhibitors in BGC-823 cells. (e) MiRNA-324-5p activated Wnt/β-catenin downstream gene expression. Cell lysates were used to blot Wnt downstream gene antibody 48 h after transfection of miRNA-324-5p inhibitors or mimics in BGC-823 and HFE-145 cells, respectively. (f) MiRNA-324-5p induced EMT. Cell lysates were used to blot EMT markers 48 h after transfection of miRNA-324-5p inhibitors or mimics in BGC-823 and HFE-145 cells, respectively. Each experiment was repeated three times