Fig. 2

The effect of ERβ overexpression and calycosin on TGF-β1-induced LX-2 cells proliferation. Cell counting kit-8 (CCK-8) assay was used to select the safe concentration range of calycosin on normal LX-2 cells (A) and the effective concentration range of calycosin on TGF-β1-induced LX-2 cells (B). C LX-2 cells and transfected cells (MOI = 40) were cultured for 24 h, 48 h and 72 h to show the effect of ERβ overexpression alone on cell proliferation. LX-2 cells transfected for 72 h were treated with TGF-β1 (10 ng/ml) for another 12 h to investigate the effect of TGF-β1 on proliferation of ERβ overexpressing cells. D The effect of ERβ, calycosin (50, 100, 200 μM) and E2 (1μM) on proliferation of TGF-β1-induced LX-2 cells were determined in the presence or absence of ERβ overexpression. In all the TGF-β1 induced cell groups, cells were pretreated with calycosin or E2 for 24 h and then stimulated with 10 ng/ml TGF-β1 for 12 h. Data were normalized in relation to the mean value of the control group and shown as mean ± SD of three independent experiments. ^ΔΔp < 0.01, ^ΔΔΔp < 0.001 vs control group; ^#p < 0.05, ^###p < 0.001 vs corresponding overexpression or vehicle group; **p < 0.01, ***p < 0.001 vs TGF-β1 group; ⁺p < 0.05, ⁺⁺⁺p < 0.001 vs the corresponding treatment groups without ERβ overexpression