Ab202419

The anti‐Ki‐67 (1:2,00, ab15580; Abcam) and anti‐SGTB (1:1000, ab202419, Abcam), biotinylated goat anti‐rabbit IgG secondary antibody (1:300, ab64256, Abcam) and 3,3‐diaminobenzidine (DAB) kits (Vector Laboratories, Peterborough, UK) were used for IHC analysis based on standard methods[18 (link)].

The total protein of each group was extracted using RIPA lysis buffer (Solarbio, Beijing, China), and determined by a BCA kit (Pierce; Rockford, IL, USA). About 30 μg proteins were subjected to SDS‐PAGE electrophoresis. After electrophoresis, the protein on the gel was transferred to the PVDF membrane (Millipore, Billerica, MA, USA) by wet transfer, and the primary antibodies againstβ‐actin (1:1000, ab8226, Abcam, Cambridge, MA, USA), cyclin D1 (1:200, ab16663, Abcam), Epithelial cadherin (E‐cadherin; 1:1000, ab231303, Abcam), Neuronal cadherin (N‐cadherin; 1:1000, ab76011, Abcam), cleaved caspase‐3 (cleaved casp‐3; 1:1000, ab2302, Abcam), and SGTB (1:1000, ab202419, Abcam) were used to incubate with the membrane overnight. Then the membrane was incubated with HRP‐conjugated secondary antibodies for 1 h. Lastly, protein bands were assessed by using Clarity™ Western ECL Substrate Kit (Bio‐Rad, Shanghai, China) and ECL system (Pierce). In each independent experiment, three parallel wells were made, and the procedures were carried out in triplicate.