Fig. 4

The effects of ERβ overexpression and calycosin on α-SMA, Col-I, MMP-1 and TIMP-1 expressions in TGF-β1-induced LX-2 cells. A Immunofluorescence images of α-SMA protein (red) in LX-2 cells with original magnification, × 400. Cell nuclei (blue) were stained with DAPI. Except cells in control and vechile groups, cells with/without ERβ overexpression were stimulated by 10 ng/ml TGF-β1 for 12 h with or without pretreatment of calycosin (200 μM) or E2 (1μM) for 24 h. (B-F) Representative blots and relative quantification analysis of α-SMA (B), Col-I (D), MMP-1 (E) and TIMP-1 (F) protein expressions detected by western blot. GAPDH was used as internal control. Data are shown as mean ± SD of three independent experiments. ^###p < 0.001 vs vehicle group; *p < 0.05, **p < 0.01, ***p < 0.001 vs TGF-β1 group; ⁺p < 0.05, ⁺⁺p < 0.01 vs the corresponding treatment groups without ERβ overexpression