Immunocytochemistry Analysis of EMT Markers

As previously explained (Yang et al., 2017 (link)), immunocytochemistry was carried out following exposure of HCT116 cells to cisatracurium. In summary, HCT116 cells were incubated in PBS containing 4% para-formaldehyde at 25°C for 20 min. For permeabilization, HCT116 cells were further maintained in PBS containing 0.5% Triton X-100 at 4°C for 10 min. After blocking using 3% BSA reagent, cells were exposed to primary antibodies for SLUG, SNAI1, E-Cadherin, and CALD1 (Proteintech). FITC conjugated secondary antibody (Invitrogen) was added and then stained with 1 μg/ml of DAPI for 60 s. Images of samples were visualized and captured using a fluorescence microscope (Olympus BX83, Japan).