Epidemiological and observational evidence suggests that obesity is related to poor Fe status. To determine interactions between obesity, dietary Fe intake and Fe status; male, weanling C57BL/6J mice were fed either high fat diets to induce obesity or a standard diet for 16 weeks. Fe concentrations of both the high fat or control diet (4.5 vs 3.8 kcal/g) were set at: 5, 50 or 500 mg Fe/kg diet. Mice fed the high fat diets had significantly higher percentage body fat (17.9%) compared to mice fed control diets (5.3%, P<0.001). Among obese mice, dietary Fe levels did not significantly influence body composition. Conversely among lean mice, mice fed the iron excessive diet had significantly less fat mass when compared to mice fed the iron deficient diet (P<0.05). Obesity and/or dietary Fe concentration did not significantly affect plasma Fe levels. ANOVA analysis showed significant effects of diet-induced obesity, dietary Fe and an interaction between both factors on liver Fe levels (P< 0.05). Obese mice had significantly lowered liver Fe levels compared to lean cohorts fed the same amount of dietary Fe (P<0.05 for all comparisons). Moreover, lean mice fed the Fe deficient diet (5 mg Fe/kg diet) had similar liver Fe levels (127 mg Fe/kg ± 0.04) compared to obese mice fed the 50 mg Fe/kg diet (132 mg Fe/kg ± 0.05). These data suggest that obesity, independent of dietary Fe intake, influences liver Fe stores.