Immunohistochemical detection kit

The pathological tissues of 205 patients were used to produce an HCC tissue microarray. After deparaffinisation and hydration, the antigen was recovered from the microarray by heating in a sodium citrate solution. Then, the microarray was incubated in 3% H₂O₂ at room temperature for 10 min and blocked in 10% goat serum for 1 h. The array was incubated with anti‐FNDC4 (1:200, Bioss) antibody at 4°C overnight. An Immunohistochemical Detection Kit (Proteintech) was used to detect the immunohistochemical reaction. After dehydration, the microarray was fixed with neutral resin. The staining intensity and area of the microarray were evaluated by three professional pathologists using CaseViewer software.

The tumor sections were first embedded in 100% xylene-dewaxed paraffin and then rehydrated with gradient ethanol. After the endogenous peroxidase was exhausted and the antigens were blocked, IHC staining was carried out by an Immunohistochemical Detection kit (Proteintech). TUNEL staining was performed using the Cell Apoptosis Detection Kit (Trevigen). The photograph was obtained with a light microscope (Olympus). Anti-PCNA (GB11010) was purchased from Servicebio (Wuhan, China).