Figure 5.

miR-103a-3p targets the 3′-UTR of DICER1 mRNA. (a) DICER1 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 DICER1 and GAPDH were determined using anti-DICER1 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) DICER1 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-DICER1 or pMIR-DICER1-mutant luciferase constructs were made according to the sequences from miRWalk database. (d) pMIR-DICER1 or pMIR-DICER1-mutant luciferase constructs were cotransfected with mimic-miR-103a-3p or mimic-miR-control into hADSCs hADSCs. Internal control for expression analysis was GUSB. Data represent mean±s.e.m. of the ratio to the value of mimic miR-cont of pMIR-DICER1 or pMIR-DICER1 mutant (n=4), *P<0.05 compared with miR-con-transfected hADSCs, ^#P<0.05 compared with anti-miR-cont-transfected hADSCs. hADSC, human adipose tissue-derived stromal cell; miR, microRNA; UTR, untranslated region.