Tmprss6 silencing prevents maternal hepcidin degradation and modulates iron metabolism during pregnancy in mice. (A) Maternal hepatic Hamp transcription was measured by plug mating wild-type (WT) C57BL6/J mice and collecting a detailed time course. Data represent fold change (2^–ddCt, in which ddCt was calculated using the unmated group as control). (B) Experimental design for C57BL6/J pregnant mice treated with either Tmprss6 siRNA (represented in green throughout) or NTC siRNA (represented with orange open circles throughout). (C) Liver Tmprss6 mRNA expression. Data represent fold change (2^–ddCt, in which ddCt was calculated using the NTC-treated unmated group as control). (D) Maternal liver Hamp mRNA expression. Data represent fold change (2^–ddCt, in which ddCt was calculated using the NTC-treated unmated group as control). (E) Serum hepcidin levels (nanograms per milliliter). (F) Maternal liver nonheme iron (micromoles per gram dry weight). (G) Maternal spleen nonheme iron (micromoles per gram dry weight). (H) Maternal serum iron (micrograms per deciliter). (I) Maternal hemoglobin concentration (grams per deciliter). (J) Maternal MCV (femtoliters). (K) Maternal liver Id1 mRNA expression. (L) Maternal liver Atoh8 mRNA expression. (M) Maternal liver Smad7 mRNA expression. Data represent fold change (2^–ddCt, in which ddCt was calculated using the NTC-treated unmated group as control). Data points depict 1 mouse per point; capped bars denote mean ± standard deviation. Statistical differences between groups were tested by 1-way analysis of variance (ANOVA) in panel A or 2-way ANOVA in panels C-M and are represented as follows: ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. ns, not significant; RNAi, RNA interference.