Fig. 5. miR-29b-3p negatively regulates CDK6 expression.

a Both Targetscan and miRDB tools showed schematic representation of putative binding site for miR-29b-3p in 3′-UTR of CDK6. b Luciferase reporter plasmids containing wild-type or mutant 3′-UTR of CDK6 was transfected with either miR-29b-3p mimics or a control miRNA into HEK293T cells. c qRT-PCR analysis of CDK6 expression in MDA-MB-231, Hs578t, SK-BR-3, and MCF-7 cells transiently transfected with either miR-29b-3p mimics or a control miRNA. d Western blot analysis detected the expression of CDK6 after knockdown or overexpression of miR-29b-3p. MDA-MB-231 and Hs578t cells were transfected with miR-29b-3p mimics or overexpression plasmid of CDK6. SK-BR-3 and MCF-7 cells were transfected with miR-29b-3p inhibitor or siCDK6 plasmid. Then e cell viability, f cell cycle, and g colony formation were performed. h IC₅₀ of palbociclib for the four breast cancer cells after transfected as indicated. Error bars indicate mean ± standard deviation.