Notch1 overexpression induces an EMT phenotype in ICC-9810 cells. (A) Increased Notch1 expression was accompanied by enhanced expression of α-SMA and Vimentin and loss of E-cadherin expression, which are hallmarks of EMT. (B) Overexpression of Notch1 results in morphological changes in ICC-9810 cells. Photographs were taken using a Nikon microscope (phase contrast). Original magnification × 400. (C and D) ICC-9810 cells were infected with Notch1 or vector for 36 h before fixation and processing for immunofluorescence staining. Actin filaments were visualized by staining with Alexa Fluor 633-Phalloidin (red). Notch1 was visualized by staining with an anti-Notch1 antibody followed by Alexa Fluor 488-conjugated anti-rabbit IgG (green). Nuclei were stained with DAPI (blue). Images were taken by confocal microscopy. Original magnification × 630.