Figure 1

Knock-down of SOX2 in 2102Ep and NTera-2 cells. The induced RNAi of SOX2 mRNA by SOX2 siRNA (siSOX2) facilitated a substantial and significant down-regulation of this gene’s expression in the EC cell lines. (A) As determined by qRT-PCR, very little SOX2 mRNA expression remains (6.5%) in siSOX2-transfected 2102Ep cells, while the remaining expression in siSOX2-transfected NTera-2 cells is higher (48.5%). SOX2 expression was compared to that in cells transfected with a scrambled, non-targeting siRNA (NC), which was set at 100%. SOX2 expression was normalised to GAPDH expression. All experiments were performed in biological triplicate. (B) As determined by Western blot, no SOX2 protein could be detected in the siSOX2-transfected cells of either cell line when compared to the non-targeting siRNA-transfected controls (NC), vehicle controls (VC) or non-transfected controls (NTC). GAPDH protein expression was measured to account for equal loading and transfer. (C) RNAi phenotypes of 2102Ep and NTera-2 cells, four days after SOX2 siRNA transfection. SOX2-silenced EC cells have a flattened, enlarged and more dispersed morphology, compared to SC-transfected cells, which are, similarly to untreated cells (not shown), small, defined and grown in dense colonies.