Phenotypic analysis of a HH mouse (Hfe^−/−) model after treatment after treatment with LNP-Tmprss6 siRNA. Six-week-old female mice were injected once and then harvested 14 days later (left side of graphs, 2-week experiment) or injected a total of 3 times over 6 weeks and then harvested 14 days after the final injection (right side of graphs, 6-week experiment) with 1 mg/kg of PBS, LNP-Luc, or LNP-Tmprss6. Total mRNA was harvested from WT or Hfe^−/− (n = 4 or 5 for each genotype) mice and Tmprss6 (A) or hepcidin (Hamp; B) mRNA was assessed by quantitative real-time PCR as in Figure 1. Ratios are expressed ± SEM. (C-F) Analysis of serum transferrin saturation (C; %), serum iron (D; μg/dL), nonheme liver iron (E) and total spleen iron (F) are depicted. (n = 5 for each genotype.) P values were calculated using the Student t test. ****P < .001; ***P < .005; **P < .01; *P < .05.