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. 2014 Dec 15;7(12):5275–5281.

Figure 2.

Figure 2

MiR-29b-3p suppressed CDK6 expression by targeting 3’UTR. A. Predicted miR-29b-3p binding sites in CDK6 3’UTR. Ba. Confirmation of the target gene of miR-29b-3p. HEK293T cells were co-transfected with miRNA control, miR-29b-3p mimic, anti-miR control or miR-29b-3p inhibitor and pGL3-CDK6 for dual-luciferase assay. PRL-TK containing Renilla luciferase was co-transfected for data normalization. Bb. Mutation analysis of the miR-29b-3p binding sites. When 4 nucleotides of these binding sites of miR-29b-3p in the 3’-UTR of CDK6 was mutated (pGL3-CDK6-Mu), the luciferase activity was significantly decreased in HEK293T cells co-transfected with miR-29b-3p mimics and pGL3-CDK6 compared with pGL3-CDK6-Mu. C. CDK6 protein level in miR-29b-3p mimic or inhibitor-treated HEK293T cells was detected by western blot.