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Fig. 1. Decreased Fpn1 protein expression in Pcm mutant placenta and embryonic liver is consistent with fetal iron deficiency, and contrasts with increased Fpn1 expression in embryonic spleen. At E16.5, placenta (A) and liver (B) show a graded decrease in Fpn1 protein expression on western blot. Approximate molecular mass: Fpn1, 68 kDa; actin, 41 kDa. (C) Western blot analysis reveals increased Fpn1 protein expression in E15.5 spleen. (D) Immunohistochemistry demonstrates a significant reduction of labyrinth cell expression of Fpn1 in E16.5 placenta from Pcm homozygotes. Original magnification 400x. (E) Immunohistochemistry reveals a widespread increase in Fpn1 expression in E15.5 Pcm mutant spleens. Original magnification 400x. (F) Analysis of non-heme iron demonstrates significant reduction of iron content in E15.5 Pcm livers, more severe in homozygotes. *, P<0.0001. (G) Semiquantitative RT-PCR of E16.5 liver indicates lower Hamp mRNA expression in Pcm mutants. Band sizes: Hamp, 171 bp; ß-actin, 250 bp.