Eclipse ti s inverted phase fluorescent microscope

SGC-7901 with or without transfection of anti-miR-30a and SGC-7901/DDP cells with or without transfection of miR-30a mimics were grown on coverslips. Then, the cells were fixed in methanol, permeabilized in 0.1% Triton X-100, and blocked in 1% BSA. To detect the expression of E-cadherin and N-cadherin, the coverslips were probed with primary antibodies against E-cadherin (1: 500, ab40772, Abcam) and N-cadherin (1: 100, ab76011, Abcam), respectively, at 4°C overnight. After the incubation, the coverslips were washed and further incubated with secondary Alexa Fluor^®555-conjugated donkey anti-rabbit IgG H&L (1: 500, ab150074, Abcam) and Alexa Fluor^®488-conjugated donkey anti-rabbit polyclonal antibody (1: 500, ab150073, Abcam), respectively, for 1 h at room temperature. Nuclei were stained with Gold Antifade Reagent with DAPI (Invitrogen). Digital immunofluorescent images were obtained using an Eclipse Ti-S inverted phase/fluorescent microscope (Nikon, Tochigi, Japan).