LA reduces iron burden in Hfe^−/− mice. Four-week-old Hfe^−/− mice were treated with intravenous LA (150 mg/kg body weight) or PBS for 4 weeks (n = 5 mice per group). (A) Changes in serum hepcidin levels of Hfe^−/− mice receiving LA treatment. (B-C) Changes in serum (B) and hepatic tissue (C) iron contents in Hfe^−/− mice with LA treatment. (D) Perls Prussian blue staining of liver sections (×100) from Hfe^−/− mice after LA treatment, with quantitative analysis of iron content in the right panels. (E) Immunofluorescent staining of FPN (in green), and 4′,6-diamidino-2-phenylindole (DAPI, blue) in the duodenum sections (×400) from Hfe^−/− mice upon LA treatment with quantitative analysis in the right panel. (F) Enhanced 3,3'-diaminobenzidine (DAB) iron staining of duodenum sections (×200) from Hfe^−/− mice after LA treatment, with quantitative analysis of iron content in the right panels. (G) Western blotting of Smad1, P-Smad1/5/8, CREB1, P-CREB1, FPN, and FTL in liver specimens from LA-treated mice relative to untreated control with quantitative analysis in the right panel. ∗P < .05 and ^#P < .001, relative to untreated WT mice or as indicated. Data are represented as mean ± standard deviation. WT, wild type.