Evidence for Hh pathway activation and EMT in clonally derived MF-HSC from livers with CCl4-induced cirrhosis. Clonally derived HSC lines (8B and 5H) from a single rat with CCl4-induced cirrhosis were compared with freshly isolated primary rat Q-HSC to assess markers associated with EMT and Hh signaling. A: mesenchymal marker expression (α-SMA, Col1α1, fibronectin, vimentin, and S100A4). B: Hh signaling (Shh and Gli2). C: epithelial markers (K7, desmoplakin, and E-cadherin). D: EMT markers (BMP-7 and Id2). For each gene, expression was normalized to that of a housekeeping gene (S9) in the same RNA sample and then expressed relative to expression of the same gene in primary Q-HSC. 8B were then treated with tomatidine or cyclopamine, and effects on gene expression changes were assessed by qRT-PCR. E: EMT-inhibitory markers (BMP-7 and desmoplakin) and the Q-HSC marker PPARγ. F: myofibroblastic markers (α-SMA, Col1α1, and S100A4). Values are means ± SE of triplicate experiments. *P < 0.05; **P < 0.01.