Ab207608

The tumor Sects. (4 μm paraffin embedded) were performed the dewaxing and rehydration. After sealing, the sections and primary antibody Ki67 (ab15580, 5 µg/ mL, Abcam, Shanghai, China), E-cadherin (ab231303, 1 µg/mL, Abcam), or N-cadherin (ab207608, 1/500, Abcam) were blended for 12-h incubation at 4 °C and further mixing for appropriate secondary antibody (ab6721, 1/1000, Abcam, Shanghai, China). Next, the sections were made the dyeing by diaminobenzidine (DAB) and re-dyeing by hematoxylin. Lastly, images were obtained under a microscope (Nikon, Tokyo, Japan).

Cells and tissues were lysed using RIPA lysis buffer (Beyotime) to obtain protein samples. After the protein concentration was measured using a BCA kit (Beyotime), the corresponding volume of protein was supplemented with sample loading buffer (Beyotime), mixed, and heated in boiling water for 3 min to denature the protein. Electrophoresis was performed at 80 V for 30 min and 120 V for 1-2 h after the addition of bromophenol blue for the separation gel. The protein was transferred to a membrane in an ice bath with a current of 300 mA for 60 min, after which the membrane was rinsed for 1-2 min and sealed for 60 min or blocked overnight at 4°C. Primary rabbit antihuman GAPDH (ab9485, 1:5000, Abcam, Cambridge, UK), rabbit antihuman LSM4 (ab236744, 1:500, Abcam), mouse antihuman E-cadherin (ab231303, 1:1000, Abcam), rabbit antihuman N-cadherin (ab207608, 1:1000, Abcam), and mouse antihuman vimentin (ab8978, 1:1000, Abcam) were incubated on a shaker for 2 h, followed by three rounds of washing (each for 10 min). Next, a secondary antibody (goat anti-rabbit or goat anti-mouse IgG tagged with horseradish peroxidase, 1:5000, Beijing ComWin Biotech Co., Ltd., Beijing, China) was added for 1 h incubation. After color development, a chemiluminescence imaging system (Bio-Rad, Hercules, CA, USA) was used for detection.