SnPP injections decrease nonheme iron levels and heme-iron recycling in the liver of Th3/⁺mice. (A) Representative pictures (20× magnification; scale bar, 200 µM) of Perls’ Prussian blue staining in Wt and Th3/⁺ mice injected with either PBS or SnPP. (B) Ferrozine assay of nonheme iron content in the liver isolated from mice treated as follows: Wt + PBS (n = 6), Wt + SnPP (n = 6), Th3/⁺ + PBS (n = 5), or Th3/⁺ + SnPP (n = 5). (C) qRT-PCR analysis of HAMP mRNA expression in the liver of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 3). (D) Enzyme-linked immunosorbent assay of hepcidin in the serum of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 6). (E) Western blot analysis of FPN protein expression in samples isolated from the liver of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 3). (F) Representative images (20× magnification; scale bar, 200 µM) of immunohistochemical staining of FPN in livers from Wt or Th3/⁺ mice injected with PBS or SnPP. (G) Percentage of ⁵⁹Fe in the liver of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 3). (H) Percentage of ⁵⁹Fe in the heme fraction in the liver of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 3) (I) Percentage of ⁵⁹Fe in total peripheral blood of Wt and Th3/⁺ mice injected with either PBS or SnPP (n = 3). Percentage was calculated based on the total ⁵⁹Fe recovered for each mouse. *P < .05, ***P < .001.