Figure 4. Nintedanib inhibits the expression of α smooth muscle actin (α-SMA) and Snail induced by transforming growth factor β1 (TGF-β1). A: The cells were treated with TGF-β1 (5 ng/ml) in the presence or absence of SB203580 or nintedanib (1 μM) for 24 h and then stained with anti-α-SMA linked to fluorescein isothiocyanate (FITC) and 4’,6-diamidino-2-phenylindole (DAPI; nuclei). Immunofluorescence was detected by confocal microscopy. Scale bar = 200 μm. B,C: Human Tenon’s fibroblasts (HTFs) were treated with TGF-β1 (5 ng/ml) in the presence or absence of SB203580 or nintedanib (1 μM) for 24 h. The mRNA (B) and protein (C) expressions of Snail were analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively, and normalized to GAPDH expression. Data are presented as the means ± standard deviations (SDs; n = 3) of independent repeated experiments (* p<0.01;** p<0.01;*** p<0.001; **** p<0.0001 compared with the control or TGF-β1 group).