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. 2015 Jul 10;47(7):e172. doi: 10.1038/emm.2015.39

Figure 3.

Figure 3

miR-103a-3p targets the 3′-UTR of CDK6 mRNA. (a) CDK6 expression in hADSCs transfected with oligonucleotides was analyzed by western blot. Lysates were prepared at 2 days after transfection with oligonucleotides. To confirm equal loading, the quantities of CDK6 and GAPDH were determined using anti-CDK6 and anti-GAPDH antibodies. The relative expression ratio of each protein was quantified by densitometric evaluation of western blots. Data represent mean±s.e.m. (n=4). *P<0.05 compared with mimic-miR-cont transfected hADSCs. (b) CDK6 expression in hADSCs transfected with oligonucleotides was analyzed by real-time PCR. Total RNAs were isolated at 2 days after transfection with oligonucleotides. Data represent mean±s.e.m. (n=4). *P<0.05 compared with mimic-miR-cont transfected hADSCs. (c) pMIR-CDK6 or pMIR-CDK6-mutant luciferase constructs were made according to the sequences from miRWalk database. (d) These contructs were cotransfected with mimic-miR-103a-3p or mimic-miR-control into hADSC. Data represent mean±s.e.m. of the ratio to the value of mimic miR-cont of pMIR-CDK6 or pMIR-CDK6 mutant (n=4), *P<0.05 compared with mimic miR-cont-transfected hADSCs. hADSC, human adipose tissue-derived stromal cell; miR, microRNA; UTR, untranslated region.