Figure 8

Activation of α7 nAChR enhances TGF-β signaling. (A) Flow cytometry analysis of α7 nAChR expression in fibroblasts. The cells were collected and stained with fluor-633 α-BTX at 0, 0.25, 0.5 and 1 µg/mL. (B, C) Effect of GTS-21 on Acta2 in BLM-challenged fibroblasts and lung epithelial cells. The cells were pretreated with GTS-21 and then stimulated with or without BLM (10 µM). (B) Changes of Acta2 mRNA in fibroblasts at 24 h. (C) Acta2 mRNA expression in A549 cells at 24 h. *p < 0.05 for control and BLM-stimulated groups. N = 3 in each group. (D, E) Effect of GTS-21 on Acta2 in rhTGF-β1-challenged lung epithelial cells and fibroblasts. (D) Changes of Acta2 mRNA in A549 cells at 24 h. (E) Changes of Acta2 mRNA in fibroblasts at 24 h. The cells were pretreated with GTS-21 (25, 50 µM) and then stimulated with or without rhTGF-β1 (5 ng/mL). **p < 0.01 for rhTGF-β1 versus rhTGF-β1 + GTS-21 groups. N = 3 in each group. Data are mean ± SD. Experiments were repeated three times. (F) Immunoblotting analysis of activation of α7 nAChR on rhTGF-β1-induced fibrogenic signaling. The cells were pretreated with GTS-21 (25 µM) and then stimulated with or without rhTGF-β1 (5 ng/mL). The cells were harvested after rhTGF-β1 stimulation for detecting p-PTP1B, p-Smad2/3 and µ-SMA. Experiments were repeated three times.