4 6 diamidino 2 phenylindole dapi zli 9557

An equivalent amount of SW480 cells (n=5×10⁵) was seeded on sterilized glass coverslips. Briefly, after fixation with methanol for 20 min and blocking with goat serum (1:10 dilution; cat. no. ZLI-9022; ZSGB-BIO) for 1 h at room temperature, the coverslips were incubated with primary antibodies against vimentin, N-cadherin and E-cadherin (1:100 dilution; product nos. 5741, 13116 and 14472, respectively; Cell Signalling Technology) overnight at 4°C, followed by Alexa 488-conjugated goat anti-rabbit antibody and Alexa 555-conjugated goat anti-mouse antibody (1:100 dilution; product codes A11008 and A21422, respectively; Invitrogen; Thermo Fisher Scientific, Inc.) for 1 h at room temperature. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; ZLI-9557; ZSGB-BIO) for 30 min at room temperature. The fluorescent expression of the target markers and the nuclei were evaluated and imaged using a Leica confocal laser microscope (magnification, ×400).