FIGURE 5.

CKI suppresses HSCs activation by restraining TGF‐β/Smad signaling in vitro. (A) LX‐2 cells were treated with 5 ng/ml TGF‐β1 along with 1 mg/ml CKI for 6, 12, and 24 h. mRNA expressions of TGF‐β1, COL1A, Fibronectin, and TIMP1 were detected by qRT‐PCR in LX‐2 cell lysates. (B) LX‐2 cells were treated with or without 5 ng/ml TGF‐β1 along with different dosages of CKI (0.5, 1, and 1.5 mg/ml) for 12 h. mRNA expressions of TGF‐β1, COL1A, Fibronectin, and TIMP1 were detected by qRT‐PCR in LX‐2 cell lysates. (C) Western blot for TGFβ, Fibronectin, and p‐Smad2/3 in LX‐2 cells from A. (D) Western blot for TGFβ, COL1A, Fibronectin, and TIMP1 protein levels in LX‐2 cells from B. (E and F) qRT‐PCR analysis of HSCs activation markers αSMA and desmin mRNA expression in LX‐2 cells. (G) Protein expressions of αSMA and desmin were quantified by Western blot in LX‐2 cells. (H) Western blot analysis of TGFβR1, TGFβR2, p‐Smad2, total Smad2, p‐Smad3, total Smad3, Smad4, Smad7, and Smurf2 in LX‐2 cells. Data are presented as means ± SEM. ns, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001