Primary antibody against e cadherin

Cells were seeded on a millicell slide (Millipore, Billerica, MA, USA) and incubated until cell stretched. Paraformaldehyde (4%) was used to fixate cell and then Trition‐X100 (0.3%) used to increase the membrane permeability. After blocking by goat serum, the slide was incubated in primary antibody against E‐cadherin, N‐cadherin, Snail, vimentin, β‐catenin (Cell Signaling Technology, Beverly, MA, USA) at 4° overnight. Next, slide was incubated with fluorochrome‐labeled secondary antibody (Alexa Fluor 488; life technologies, USA) at room temperature and for 30 minutes. DAPI was used to stain nucleus. Finally, observing the result of staining and getting pictures under confocal laser‐scanning microscopy (FluoView FV10i; Olympus, Japan).

The protein in TNBC cells was extracted using cell lysis buffer (St. Louis, MO). There was 20-30 μg protein resolved on 10% SDS-PAGE gels and transferred onto PVDF membrane (Millipore, Merck, United States). We incubated sliced membranes and primary antibody against E-cadherin, N-cadherin, Vimentin, Bax, Bcl-2, GADD45α, GAPDH, and β-actin (Cell Signaling, Danvers, MA) overnight at 4°C. β-actin or GAPDH was selected as the loading control. After incubating sliced membranes and secondary antibody for 1 h, the detection was conducted via ECL Advance Reagent (Millipore, Merck, United States) and Image Lab Software (Bio-rad, Kidlington, UK).