Fig 5. TDO mutation of DLX3 inhibits the activation of EMP genes by wild-type DLX3.

(A) The plasmids pCI-neo (control empty vector, con), V5DLX3^WT, and V5DLX3^TDO were transfected into LS8 cells. Equal amounts of V5DLX3^WT or V5DLX3^TDO plasmid were added to each group, and the total amounts of plasmid in the groups were kept constant using pCI-neo. Over-expression of V5DLX3^WT or/and V5DLX3^TDO were analyzed by western blot using antibody against V5-tag. A representative figure from three independent experiments is shown. (B) After over-expression, the mRNA expression levels of Enam, Amelx, Klk4, and Odam were evaluated. Values were obtained from three independent experiments, and are presented as mean ± SD. *P <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. (C) Left panel: western blot for the protein expression levels of ENAM, AMELX, and KLK4 after transfection (representative of three independent experiments). Right panel: densitometric analysis of images of 3 independent experiments. *P <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. (D) After co-transfection with pCI-neo (empty control vector), V5DLX3^WT, V5DLX3^TDO, or both V5DLX3^WT and V5DLX3^TDO plasmids, the relative transcriptional activity of the 5 reporter constructs containing potential DLX3 response elements were analyzed by luciferase assays. Data were compared with the control group (con), and are presented as mean% ± SD. *P <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. pEnam-E1 represents pGLEnam-E1, etc.