Apo-Tf injection exacerbates responses to dietary iron loading and promotes NTBI formation in mice fed a high-iron diet. (A) Schematic experimental outline. Male 7- to 8-week-old Tfrcfl/fl mice (n = 6-10 per group) were fed a standard diet (200 ppm iron), iron-deficient diet (IDD; 2-6 ppm iron), or HID (2% or 0.25% carbonyl iron), as described by Charlebois et al.2 Where indicated, the mice were injected intravenously (tail vein) with 10 mg apo-Tf. At the end point, the animals were sacrificed; serum was prepared, and livers were harvested for biochemical analysis. (B) Serum TBI. (C) Transferrin saturation. (D) Total iron-binding capacity (TIBC). (E) NTBI. (F-I) Quantitative polymerase chain reaction analysis of liver Hamp, Bmp6, Bmp2, and Id1 mRNAs. Serum data (B-E) are represented as mean ± SEM; gene expression data (F-I) are represented as geometric mean ± geometric SD. Statistical differences (P < .05) were determined using 1-way analysis of variance. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001.
