Fig. 2

FRA affected collagen metabolism and HSFs activation in vitro. (A) qRT‒PCR results of the mRNA levels of COL1A1, COL1A2 and COL3A1 in HSFs after treatment with FRA (100 µM, 200 µM and 300 µM) for 48 h. GAPDH served as the control. (B) The protein levels of COL 1 and COL 3 in HSFs incubated with FRA for 48 h were determined by WB analysis. (C) Quantification of COL 1 and COL 3 expression based on WB analysis. (D) Immunofluorescence staining for COL1A1 and COL3 in HSFs after treatment with FRA for 48 h (scale bar = 50 μm). (E) The statistical results of the relative fluorescence intensity of COL1A1 and COL3. (F) α-SMA mRNA expression in HSFs after incubation with TGF-β1 (5 ng/ml) and different concentrations of FRA (0 µM, 100 µM, 200 µM and 300 µM) for 48 h was determined by qRT‒PCR. (G) WB analysis of α-SMA expression in HSFs subjected to the indicated treatments for 48 h. (H) Quantification of the α-SMA protein level based on WB analysis. (I) Immunofluorescence staining for α-SMA and F-actin in HSFs treated with TGF-β1 (5 ng/ml) and FRA (0 µM or 300 µM) for 48 h (scale bar = 50 μm). (J) Quantification of the relative fluorescence intensity of α-SMA. (K) Representative images of the collagen gel contraction assay in different treatment groups on day 3. The dashed white lines indicate the areas of collagen gel. (L) Quantitative results of the collagen gel contraction assay. The data are presented as means ± SDs (n = 3 independent experiments). **P < 0.01, ***P < 0.001