Neo1^fl/fl;Alb-Cre⁺mice have iron overload. (A) Serum iron (Fe) assay for Neo1^fl/fl;Alb-Cre⁻, Neo1^fl/wt;Alb-Cr⁺, and Neo1^fl/fl;Alb-Cre⁺ male and female mice at 5, 8, and 15 weeks old. (B) Liver nonheme iron assay. Iron concentration is expressed as micrograms of iron per gram of wet tissue. (C) Representative images of western blot analysis for Neo1, Tfr2, Tfr1, and β-actin in the liver extracts of 8-week-old Neo1^fl/fl;Alb-Cre⁻, Neo1^fl/wt;Alb-Cr⁺, and Neo1^fl/fl;Alb-Cre⁺ mice. (D) Spleen nonheme iron assay. Each group consists of at least 3 animals. The means ± SD are presented. A 2-tailed Student t test was used to analyze the data for 5-week-old mice, and 1-way analysis of variance (ANOVA) and Tukey’s posttests were used to analyze the data for 8- and 15-week-old mice. *P < .05; **P < .01; ***P < .001.