The Role of TWIST in Angiogenesis and Cell Migration in Giant Cell Tumor of Bone
Figure 3
The effect of TWIST siRNA on E-cadherin expression in the mesenchymal stromal cells of GCT. (a) The expression of TWIST in untreated (CT) and TWIST overexpressed (GCTT) GCT cells was determined using qRT-PCR in triplicate. TWIST overexpression resulted in a statistically significant decrease in E-cadherin expression and increase in fibronectin and MMP-2 expression at the transcript level (). (b) Validation of transcript data by western blots. (c) The expression of empty vector treated (UT) and TWIST knockdown (GCT KO) GCT cells was determined using qRT-PCR in triplicate. TWIST knockdown resulted in a statistically significant increase in E-cadherin expression in GCT cells (). (d) Western blot demonstrated the same trend in TWIST and E-cadherin protein levels in GCT stromal cells transfected with TWIST siRNA compared to untransfected GCT cells. (e) Subcellular localization of E-cadherin in TWIST-depleted and GCT untransfected stromal cells was detected by immunofluorescence using an anti-E-cadherin antibody. DAPI staining (blue) indicates the nuclei, and phalloidin (red) is used for cytoskeleton staining (oil immersion; 100x magnification).
