Fig. 3

DHM inhibited HSCs activation by inducing autophagy. a The protein levels of LC3B-II and SQSTM1 in the mouse liver were analyzed by western blot. b–c The quantification of LC3B-II (b), and SQSTM1 (c) were displayed by histogram, respectively. Data were presented as mean ± SEM (n = 3); d HSCs were treated with DHM (30 μM) for 2 h, then cells were exposed to TGF-β1 (5 ng/mL) for an additional 24 h. BafA1 (10 nM) was added 1 h before DHM treatment. The expressions of LC3B-II and SQSTM1 were detected by western blot. e–f Bar charts show the quantification of endogenous LC3B-II (e) and SQSTM1 (f). g LX2 cells were pretreated with DHM (30 μM) in the presence or absence of 3-MA (5 mM) for 2 h, followed by treatment with TGF-β1 (5 ng/mL) for another 24 h. Representative (TEM) images of LX2 cells after treatment. Red arrows indicate the autophagosomes. h–i Cells were treated as described in (G), and the levels of collagen I (h) and α-SMA (i) in the medium were detected via ELISA assay. Data were presented as mean ± SEM (n = 3); *p < 0.05, **p < 0.01, compared between the marked groups. 3-MA (3-methyladenine); BafA1, bafilomycin A1; LC3B-II, microtubule associated protein 1 light chain 3 beta; SQSTM1, sequestosome 1; TEM, transmission electron microscopy; TGF-β1, transforming growth factor-beta 1