The regulation of E-cadherin expression by Twist2 in breast cancer cells.

<p><b>A.</b> Immunoblot analysis showing that strong expression of E-cadherin was found with cytoplasic Twist2 expression. Stable expression of ectopic Twist2 in breast cancer cells (Twist2/MCF-7) were verified by western blot with anti- Twist2 and anti-flag antibodies. No obvious changes of E-cadherin was detected between Twist2/MCF-7, Vec/MCF-7 (the vector control), and the parental group. <b>B.</b> Immunoblot analysis of Twist2 in subcellular fractions showing that Twist2 was localized in the cytoplasm. Oct-1 indicated nuclear fraction and IkB-α indicated cytoplasmic fraction. The cells were from the stably transfected samples. <b>C.</b> Immunofluorescent staining of Twist2 and E-cadherin in MCF-7 cells showing cells with Twist2 (in red) in cytoplasm expressed E-cadherin (in green) on cell membrane. Nuclei were counterstained with DAPI (in blue). The cells were from the stably transfected samples. <b>D.</b> Immunofluorescent staining showing that transient over-expression of Twist2 (in red) in nuclei caused loss of E-cadherin in the same cancer cells. Cells without nuclear Twist2 retained expression of E-cadherin on membrane. Nuclei were counterstained with DAPI (in blue).