Fig. 4. Ser 68 phosphorylation is required for Ras-stimulated and Twist1-promoted breast cancer cell invasion.
A. Stable MCF-10A cell lines with mock control, Twist1 expression or Twist1-S68A expression were assayed by immunoblotting (IB) with antibodies indicated. Twist1, S68A-Twist1 and GAPDH mRNAs were measured by RT-PCR. B. The above cells were immunostained by the E-cadherin antibody (green color in all panels). Cell nuclei were stained by DAPI (right panels). C. Morphologies of MCF-10A cell lines expressing H-RasV12 and Twist1, S68A-Twist1 or a mock control as indicated (upper panel). Real-time cell invasion assays (lower panel) were performed with the 3 cell lines in the upper panel and the 3 cell lines in panel A.