Fig. 3

Avitinib inhibits fibroblasts activation via TGF-β1/Smad signaling pathway in NIH-3T3 cells. AVB (1.25, 2.5 µM) and/or TGF-β1 (5 ng/mL) was exposed to NIH-3T3 cells for 24 h, and the α-SMA expression level was detected by cellular immunofluorescence. Scale bar:50 µm;(B-C) AVB (1.25, 2.5 µM) and/or TGF- β1 (5 ng/mL) was exposed to NIH-3T3 cells for 12 h, and quantitative real-time PCR was used to analyze the mRNA transcription level of Col 1 and α-SMA. D-F TGF-β1 (5 ng/mL) and/or AVB (1.25,2.5 µM) were exposed to NIH-3T3 cells for 24 h, and the Col 1 and α-SMA expression levels were detected by Western Blot. Quantitative analyses of Western blot are shown beside. G-I NIH-3T3 cells were exposed to TGF- β1 (5 ng/mL) for 0.5 h and AVB (1.25, 2.5 µM) for 2 h, and detecting the protein expression levels of Smad3, Smad2, p-Smad2 (S467) and p-Smad3 (S423/425) by Western blot. Densitometric analyses are shown beside. Data was presented as the means ± SD, n = 3. ^##P < 0.01, ^####P < 0.0001 versus control group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus TGF-β1 group