Knock down of Dact2 activates endogenous Pitx2 target genes.

(A) Knocking down of endogenous Dact2 in LS-8 cells by shRNA was shown by Western Blot. Negative control shRNA transfected cells show no change. GAPDH was probed as loading controls. Protein band intensities were quantified and shown as relative value ±SEM. (B) mRNAs were extracted from LS-8 cells transfected with Dact2 shRNA or NC shRNA, and subjected to RT-PCRs and real-time PCRs. Relative expression levels of Dlx2 and Amelx were analyzed and correlated with Dact2 expression level. All Real-time PCRs were performed in triplicates and repeated six times.