Effect of inflammation on Smad1/5/8 phosphorylation and liver iron in wild-type C57BL/6 and Hfe KO mice. (A) Western blotting of liver lysates from saline- or LPS-injected WT and Hfe KO mice using an antibody to phosphorylated Smad1/5/8; β-actin was used as loading control. (B) Chemiluminescence was quantified using Quantity One software to calculate the ratio of phosphorylated Smad1/5/8 to β-actin. Mean ratios of four samples (± SEM) are represented relative to the mean ratio of the saline-injected WT or Hfe KO mice. C) Liver iron was quantified; iron levels are expressed as μg/g dry weight. Data are mean ± SEM (n = 5).* denotes significant difference from LPS-injected WT mice (p <0.05). ** denotes significant difference from the saline–injected WT mice (p <0.01).