TMAs were purchased from Shanghai Biochip Co., Ltd., (Shanghai, China). The histopathological diagnosis was determined by the World Health Organization criteria. Tumor staging was defined according to the 6th edition of the tumor-node-metastasis (TNM) classification of the International Union Against Cancer. Overall survival (OS) was termed as the interval between the dates of surgery and death or final follow-up.
Immunohistochemistry staining of paraffin sections were manufactured on 4 µm-thick slides, followed by EnVision two-step procedure of Dako REAL™ EnVision™ detection system (Dako, Carpinteria, CA, USA). After antigen retrieval, the slice was incubated with the primary antibodies USP20 (1:250; Proteintech Group, Chicago, IL, USA) and Claspin (1:1500; Proteintech Group) overnight at 4°C, followed by incubation with the HRP labeled secondary antibodies at 37°C for 0.5 h and slide was visualized by diaminobenzidine.
Immunohistochemistry staining of paraffin sections were manufactured on 4 µm-thick slides, followed by EnVision two-step procedure of Dako REAL™ EnVision™ detection system (Dako, Carpinteria, CA, USA). After antigen retrieval, the slice was incubated with the primary antibodies USP20 (1:250; Proteintech Group, Chicago, IL, USA) and Claspin (1:1500; Proteintech Group) overnight at 4°C, followed by incubation with the HRP labeled secondary antibodies at 37°C for 0.5 h and slide was visualized by diaminobenzidine.