Figure 5. Sp1 is a direct target of miR-199a-3p. (A) The potential binding sites of miR-199a-3p in Sp1 mRNA were predicted by TargetScan. (B) HEK-293T cells were co-transfected with the miR-199a-3p mimics or inhibitor and Sp1-WT or Sp1-MUT luciferase reporter plasmids. The cells were harvested 48 h after transfection for luciferase activity determination. (C) qPCR results showed that the upregulating effect of MPP+ on Sp1 mRNA expression was inhibited by miR-199a-3p overexpression. (D, E) Western blot analysis was performed to measure the protein expression of Sp1 upon the miR-199a-3p transfection. The data are representative of three experiments. *p <0.05, **p <0.01 and ***p <0.001.