Effects of hepcidin-mediated maternal iron restriction on the placenta. Placentas from the 3 minihepcidin treatment groups from Figure 4 were analyzed at E18.5 for weight and iron content. (A) Placental weight was used as a surrogate measure for placental development. (B) Placental nonheme iron content. (C) Western blot of total placental TFR1 and FPN protein for the 50-6d and 50-11d treatment groups (left panel) and 10-11d treatment group (right panel). Quantification of western blot results for TFR1 (D) and FPN (E) protein relative to GAPDH. (F) Calculated PIDI, the ratio of placental FPN to TFR1 protein. Statistical comparisons were performed by 2-tailed Student t test for normally distributed values, 2-tailed Mann-Whitney rank-sum test for nonnormally distributed values (indicated by * following P-values), or 2-tailed t test with Welch’s correction for normally distributed but nonequal variance data sets (indicated by ^# after the P value).