Validation of CDK6 and TAB2 as target genes for miR-29a and miR-142-3p, respectively. (A) The predicted miR-29a binding sites in the 3′-UTR of CDK6 mRNA are shown. CDK6_WT is the pMIR-REPORT construct containing the entire 3′-UTR sequence of CDK. CDK6_MUT1, MUT2, and DM are the pMIR-REPORT constructs containing mutated nucleotides in the first, second, and both miR-29a binding sites, respectively. (B) Luciferase reporter assays. Relative luciferase levels in 293T cells cotransfected with the CDK6_WT, CDK6_MUT1/2, or CDK6_DM report construct and either miR-29a mimic or scrambled control. *P < .05 (Student t test). **P < .01 (Student t test). (C) Western blot analysis of CDK6 levels in 293T, THP-1, and NB4 cells. miR-29a mimic, inhibitor (anti-29a), or a control was transfected into the indicated cells. A representative Western blot from 3 independent experiments is shown. (D) The predicted miR-142-3p binding site in the 3′-UTR of TAB2 mRNA is shown. TAB2_WT is the reporter construct containing the entire 3′-UTR sequence of TAB2, and TAB2_MUT is the reporter construct containing mutated nucleotides at the miR-142-3p binding site. The mutated nucleotides of the 3′-UTR site are indicated by gray boxes. (E) Relative luciferase expression levels in 293T cells cotransfected with TAB2_WT or TAB2_MUT with either miR-142-3p mimic or a scrambled control. (F) Western blot analysis of TAB2 levels in 293T, THP-1, and NB4 cells. miR-142-3p mimic, a miR-142-3p inhibitor (anti–142-3p), or a control was transfected into the indicated cells. A representative Western blot from 3 independent experiments is shown.