Fig. 5

TNF-α mediates the ITGA5 elevation and NF-κB pathway activation. IPF/normal supernatants were added to fibroblast cells seeded on plastic or fibronectin (FN) (10 μg/ml) coated plates for 30 min. Following culture, cells were harvested for protein extraction and the effect of IPF derived supernatants (IPF-SN) on pIκBα (a) and pSTAT3- Tyr705 and pSTAT3- ser727 (b) levels were tested by western blotting (n = 4). Infliximab (200–400 μg/ml) was added to fibroblasts cultured with normal (N-SN)/IPF-SN. Levels of ITGA5 were analyzed by western blotting following 24 h (n = 8 and n = 4 for 200 and 400 μg/ml, respectively) (c) and of pIκBα (n = 4) (d) and pSTAT3-Tyr705 (n = 4) (e) following 30 min. Results were normalized to control (N-SN) and considered significant * if p < 0.05, error bar represents the standard error