(A) Conditioned medium from WT and Smo KO hepatocytes was collected and analyzed to determine the concentration of osteopontin (OPN) by ELISA. (B) Western blot and qRT-PCR analysis of OPN in hepatocytes transfected with Gli1 or control vector for 48 h. (C, D) Relative OPN mRNA and protein expression in WT and Smo KO hepatocytes treated with vehicle, 0.5 μM SAG or 300 ng/ml Shh ligand, as analyzed by qRT-PCR and Western blotting, respectively. (E) WT hepatocytes were treated with vehicle, 0.5 μM SAG or 0.5 μM SAG plus 5 μM GANT61 for 48 h and the expression of OPN was determined by western blotting. The data are expressed as mean ± SD (***P<0.001).