Itgb8

Total protein was extracted from cells using lysis buffer, followed by determination of total protein concentration in the sample. Each protein sample (50 µl) was separated by 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and then transferred to polyvinylidene difluoride membranes. Membranes were blocked using 5% non-fat milk for 30 min at room temperature, then incubated with primary antibodies (1:1000 dilution; anti-ANGPT2, CD19, COL4A3, FGF18, ITGB4, ITGB8, LAMA3, LAMC2, PPP2R2C, SGK2, SYK, AKT3, COL6A1, CSF3, FGF1, ITGA2, ITGA11, MYB, PCK2, PGF, PIK3AP1, SGK1, TLR4 and TP53 (all Abcam, Cambridge, UK) at 4 °C overnight. Membranes were washed three times using 1 × Tris-buffered saline-Tween-20 (TBST), and then incubated with the corresponding horseradish peroxidase (HRP)-conjugated secondary antibody (1:5000 dilution; all Abcam) for 1 h at 37 °C, and again washed three times with 1 × TBST. GAPDH was quantified as the loading control. The blots were visualized using enhanced chemiluminescence (ECL) reagent (Cwbiotech, Beijing, China) and Image Lab software, version 5.1 (Bio-Rad Laboratories, Hercules, CA, USA).