Research Paper Volume 12, Issue 17 pp 16936—16950

Figure 4. CDK6 is a target of miR-34a-5p in nasopharyngeal cancer cells. The relative CDK6 level (fold) in CNE-2R and 6-10BR cells versus CNE-2 and 6-10B cells measured by miR-omic shown in a table (A), qRT-PCR shown in a plot (B) and western analyses shown in (C). “-” indicates no detection in the omic analysis. CDK6 protein levels determined western blot analyses in the miR-34a-5p antagomiR (5PA) -transfected CNE-2R and 6-10BR cells (D) and the miR-34a-5p mimic (5PM)-transfected CNE-2 and 6-10B cells (E) versus the negative control (NC). (F) Luciferase reporter constructs: WT and MUT CDK6 in the miR-34a-5p-binding sites were inserted into psiCHECK-2 vector. The blue region is the binding site. The CDK6 site is predicted to be a target of miR-34a-5p. One seed sequence mutant of miR-34a-5p was shown below. The relative luciferase activities (fold) of the reporter with the wild-type (WT) or mutant-type (MUT) CDK6-UTR or without the UTR (NC) were determined in the nasopharyngeal cancer cells transfected with the miR-34a-5p mimic (in CNE-2R) (G) or antagomiR (in CNE-2) (H).