Effect of osteopontin (OPN) and tumor necrosis factor-α receptor (TNFR1) deficiency on the degree of α-SMA expression in DDC-fed mice. Immunohistochemistry for α-SMA (a–d) and α-SMA protein measurement by western blotting (e) in chow-fed wild type (WT/chow), WT 4 weeks DDC-fed (WT/DDC), OPN knock-out 4 weeks DDC-fed (OPN/DDC) and TNFR1 knock-out 4 weeks DDC-fed (TNFR/DDC) mice. (b–d) Please note that OPN and TNFR1 loss do not reduce hepatic α-SMA expression in 4 weeks DDC-fed mice. (e) Hepatic α-SMA protein quantification by western blotting in 4 weeks DDC-fed mice revealed significant induction in all genotypes, which was more pronounced in DDC-fed OPN knock-out mice. *Indicates P<0.05 chow-fed mice vs DDC-fed mice. #Indicates P<0.05 DDC-fed OPN knock-out mice vs DDC-fed WT and DDC-fed TNFR1 knock-out mice. Ha, hepatic artery; pv, portal vein. Original magnification × 20.