Anti fndc4

The anti-FNDC4, anti-MYOG and anti-desmin were from Bioss Biotechnology. The anti-GAPDH, anti-Phospho-paxillin (Tyr118), anti-paxillin, anti-Phospho-FAK (Tyr925), anti-FAK, anti-Vinculin and anti-ITGβ1 were purchased from Cell Signaling. The anti-6× his and anti-IgG were from Santa Cruze Biotechnology. The secondary antibodies were HRP-labeled and FITC-labeled were from Beijing Biosynthesis Biotechnology Co., Ltd. PF562271 was from Selleck and used concentration for 10 nM/ml to treat cells.

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.