Gb12192

The sections of colon tissue were blocked by 3% H₂O₂ and goat serum; then, the samples were incubated with the following primary antibodies at 4°C overnight: ZO-1 (1 : 200, GB111981, Servicebio), occludin (1 : 600, GB111401, Servicebio), claudin-1 (1 : 400, GB11032, Servicebio), E-cadherin (1 : 500, GB12082, Servicebio), and vimentin (1 : 1000, GB12192, Servicebio). Then, sections were washed with PBS and incubated with the appropriate secondary antibody (Dako Real Envision/HRP, Rabbit/Mouse, K5007) at room temperature for 1 h. After that, 3,3′-diaminobenzidine with peroxidase substrate (Dako Real DAB Chromogen, K5007) were used for chromogenic reaction. Counter staining was performed with hematoxylin.

Tissue slides that were formalin-fixed and paraffin-embedded were dewaxed and rehydrated with graded ethanol and were blocked for endogenous peroxidase activity. Antigen slides were retrieved using a pressure cooker. The slides were incubated with 10% serum for 30 min at room temperature to block nonspecific antibody binding. Subsequently, the slides were incubated with anti-Ki-67 (AF1738, Beyotime; 1:100 dilution), anti-PCNA (AF0261, Beyotime; 1:200 dilution), anti-E-Cadherin (AF0138, Beyotime; 1:200 dilution), anti-N-Cadherin (22018-1-AP, Protientech; 1:2000 dilution) and anti- Vimentin (GB12192, Servicebio; 1:1000 dilution) at 4 °C overnight.