Alexa fluor 488 conjugated anti rabbit and anti mouse or anti goat secondary antibodies

Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS). Following fixation, the cells were incubated at 4°C overnight with mouse polyclonal anti-human E-cadherin (1:200) N-cadherin (1:200), anti-Jagged1 (1:200), DLL4 (1:200), Rabbit polyclonal anti-human anti-vimentin (1:200), and goat polyclonal anti-human anti-SLUG (1:200) primary antibody in PBS with 1% bovine serum albumin and 0.1% Triton X-100. Immunostaining of the proteins was visualized using Alexa Fluor 488-conjugated anti-rabbit and anti-mouse or anti-goat secondary antibodies (Molecular Probes, Seoul, Korea). Nuclei were counterstained with DAPI (Sigma-aldrich). Immunostaining was observed with an Olympus IX71 fluorescence microscope (Olympus, Seoul, Korea).