Abnormal iron distribution in Pfn2^−/−vs WT mice. Enhanced Prussian blue staining in brain (A) and liver (B) of Perls-perfused Pfn2^−/− and WT mice. In the Pfn2^−/− brain section, the arrows point to CA1-CA2 hippocampal regions, enriched in iron. In contrast, iron is lost in liver parenchymal cells. Scale is 100 µm in panel A, and 200 µm in low-magnification panels and 20 µm in the high-magnification insets in panel B. (C) Tissue non–heme iron content measured using the bathophenanthroline chromogen method in the indicated tissues shows a loss of 40% of iron in the liver of Pfn2^−/− mice. Total iron content measured by atomic absorption method in bone marrow (D) and brain areas (E) shows iron overload in olfactory bulbs, hippocampus, and midbrain of Pfn2^−/− mice. Measures are given as percentage of control WT littermates. The sample size (n) is indicated. P values were determined by unpaired 2-tailed Student t test, *P ≤ .05, **P ≤ .01.