Fig. 2.

Overexpression of IGFBPrP1 induced autophagy in primary HSCs. a Cells were cultured in 10% FBS or serum starvation medium with AdIGFBPrP1 or CAd for various time periods, and the protein levels of Beclin1, LC3B, and SQSTM1/p62 were analyzed by Western blotting. b Band intensities of Beclin1, LC3B, and SQSTM1/p62 relative to the control cells were determined after normalizing to β-actin expression. c mRNA levels of LC3B were measured by qPCR. d Cells were cultured in serum starvation medium with CAd for 6 h or AdIGFBPrP1 for 12 h. Images were acquired by TEM. Scale bar, 1 μm and 500 nm. e AVO detection after MDC staining by fluorescence microscopy. Scale bar, 20 μm. f Cells were transfected with mRFP-GFP-LC3 adenovirus and assessed using confocal microscopy. Scale bar, 50 μm. g Mean numbers of GFP and mRFP dots per cell. *P< 0.05 compared to the normal control (10% FBS). #P< 0.05 compared to the CAd control (2% FBS). Data were presented as mean ± SD for three replicate experiments (n = 3 per group)