Fig. 7

Effects of the miR-0308 mimic on CDK6 and Cyclin D1 proteins expression and the mRNA 3ʹUTR activity of CDK6 and Cyclin D1 genes. a qRT-PCR was conducted to detect the relative expression of CDK6 in both HepG2 and 7721 cells. b Western blotting analysis of CDK6 protein expression in HepG2 cells after treatment with the miR-0308 mimic for 24 h at 25 nM, 50 nM, and 100 nM, respectively. c Predicted target sites of miRNA-0308 in CDK6 3ʹUTR, and the wild-type and mutated CDK6 mRNA 3ʹUTR sequences that were inserted into the pmirGLO plasmids to construct the luciferase reporter vectors. d Activity of the luciferase reporter; higher activity indicates miR-1246 interacting with the CDK6 3ʹUTR target sequence. Cells were co-transfected with the reporter vectors and either the miR-0308 mimic or NC. Luciferase activity was analyzed at 48 h after transfection. The reporter assay was repeated three times. e qRT-PCR was conducted to detect the relative expression of Cyclin D1 in both HepG2 and 7721 cells. f Western blotting analysis of Cyclin D1 protein expression in HepG2 cells after treatment with the miR-0308-3p mimic for 24 h at 25 nM, 50 nM, and 100 nM, respectively. g Predicted target sites of miRNA-0308 in Cyclin D1 3’UTR, and the wild-type and mutated Cyclin D1 mRNA 3’UTR sequences that were inserted into the pmirGLO plasmids to construct the luciferase reporter vectors. h Activity of the luciferase reporter; higher activity indicates miR-1246 interacting with the Cyclin D1 3’UTR target sequence. Cells were co-transfected with the reporter vectors and either the miR-0308 mimic or NC. Luciferase activity was analyzed at 48 h after transfection. The reporter assay was repeated three times. *p < 0.05; n.s., no significance