Phenotypic analysis of a thalassemia-intermedia mouse (Hbb^th3/+) model after treatment with LNP-Tmprss6 siRNA. Six-week-old female mice were injected once (1 mg/kg) and then harvested 14 days later (left side of graphs, 2-week experiment) or injected a total of 3 times (1 mg/kg) over 6 weeks and then harvested 14 days after the final injection (right side of graphs, 6-week experiment). Total mRNA was harvested from WT or Hbb^th3/+ (n = 4 or 5 for each genotype) 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; g) are depicted (n = 5 or 6 for each genotype.) P values were calculated using the Student t test. ****P < .001; ***P < .005; **P < .01; *P < .05.