Fig. 2.

Mcl-1 is the downstream target gene of ssc-mir-133b during early premolar development. a In situ hybridization with DIG-labeled Mcl-1 cDNA in E40, E50, and E60 premolars. Scale bar = 200 μm. Enamel knots (EK, red dotted circle). b The Mcl-1 3′-UTR harbors one ssc-mir-133b-specific binding site. c Relative ssc-mir-133b expression levels at 48 h after transfection of ssc-mir-133b mimics and inhibitors. Relative ssc-mir-133b levels after transfection with ssc-mir-133b mimics (left). Relative ssc-mir-133b levels after transfection with ssc-mir-133b inhibitors (right). ***P < 0.001. d Relative luciferase activity is shown above. Reporter plasmids with the wild-type or mutant type Mcl-1 3’-UTR were co-transfected with ssc-mir-133b mimics or inhibitors and negative controls for both. ^#P < 0.05 compared with pMIR-REPORT-MCL1 co-transfected with the ssc-mir-133b mimic negative control and *P < 0.05 compared with pMIR-REPORT-MCL1 co-transfected with ssc-mir-133b mimics. e qPCR for Mcl-1 mRNA levels after miR-133b mimic and inhibitor transfection (left) and Western blotting of Mcl-1 protein levels after miR-133b mimic transfection (right). **P < 0.01 and ***P < 0.001