Figure 4.
Loss of function of TRβ increases Runx2 and Runx2 target genes. A, Western blot analysis illustrates that siRNA (TRβ) transfection into normal thyroid cells (ORI) results in a concentration-dependent decrease in TRβ protein and increase in Runx2 protein compared with scrambled nonspecific siRNA (ns) (upper panel). Graph summarizes quantitation, mean ± SD, of 3 separate experiments. Protein levels were standardized to β-actin and compared with respective controls. Significances at P < .01 are indicated (+). B, qRT-PCR illustrates that siRNA (TRβ) transfection results in corresponding changes in mRNA. Graph summarizes quantitation, mean ± SD of 3 separate experiments, of TRβ and Runx2 mRNA levels standardized to β-actin. Significance at P ≤ .01 compared with respective control is indicated (+). C, Knockdown of TRβ impacts Runx2 target genes; MMP13, MMP2, cyclin D1, osteopontin (OPN), and cadherin 6. qRT-PCR levels of target genes were standardized to β-actin. Data are the averaged fold change in transfected cells normalized to mock-transfected cells. Graph summarizes quantitation, mean ± SD of 3 separate experiments. Significances at P ≤ .01 compared with respective control are indicated (+).