Fig. 2

Pirfenidone and rapamycin effect in protein and gene expression of tenascin-c and fibronectin in human lung fibroblasts from IPF patients. Primary lung fibroblasts from IPF patients were treated with TGF−β (5 ng/ml), pirfenidone (1 mg/ml) and rapamycin (1 μg/ml) for 72 h. a The total cell lysates were subjected to immunoblot analysis for tenascin-c. The data shown are mean values ± SEM of four different experiments. b Results of tenascin-c transcript fold changes expressed as relative gene expression (RGE) analyzed by RT-PCR. c Fibronectin protein level. Density of protein bands were normalized against vinculin and are shown as ratios. d Fibronectin transcript fold changes expressed as relative gene expression (RGE). Charts represent mean values ± SEM of four different experiments. Levels of significance: *p < 0.05; ** p < 0.01; ***p < 0.001. * indicates the comparison between samples treated with TGF−β and all the other conditions: TGF−β versus untreated samples (control); rapamycin and pirfenidone (rapa/pirfe); and samples treated with TGF−β in combination with rapamycin (TGF−β/rapa); pirfenidone (TGF−β/pirfe) or both (TGF−β/rapa/pirfe). ^#p < 0.05; ^## p < 0.01. # indicates the comparison between untreated cells (control) and samples treated with rapamycin and pirfenidone in the absence of TGF-β (rapa/pirfe). n.s statistically not significant