Fig. 3.

Rapamycin increases IGFBPrP1-induced activation and ECM expression in primary HSCs. a HSCs were treated with rapamycin in a gradient dose and time course. Western blotting was used to analyze the expression levels of Beclin1, SQSTM1/p62, and LC3B. b Band intensities of Beclin1, SQSTM1/p62, and LC3B relative to the control cells were determined after normalization to β-actin expression. c HSCs were cultured in 10% FBS or 2% FBS with or without AdIGFBPrP1 and rapamycin (100 nmol/L) for 24 h. The protein levels of α-SMA, collagen I, IGFBPrP1, and TGFβ1 were analyzed by Western blotting. d Band intensities of α-SMA, collagen I, IGFBPrP1, and TGFβ1 relative to the control cells were determined after normalization to β-actin expression. e The mRNA levels of α-SMA, IGFBPrP1, and TGFβ1 were measured by qPCR. *P< 0.05 compared to the normal control (10% FBS). #P< 0.05 compared to the serum starvation (2% FBS). Data were presented as mean ± SD for three replicate experiments (n = 3 per group)