Excess iron upregulates Tet1 and increases 5-hmC. (A) Heatmap showing the fold change in differentially expressed epigenetic regulators in the liver of wild-type C57B/6J mice analyzed using RNA-seq data (n = 3 mice per group). (B-D) RT-PCR analysis of Tet1 mRNA measured in the indicated tissues and cells of mice fed the indicated diets (for the liver and spleen, n = 5-6 mice per group; for enterocytes, n = 5-6 mice per group; for BMDMs, n = 3-4 mice per group). (C) Fold change in 5-hmC measured in BMDMs vs FAC concentration (n = 4-5 mice per group). (D) Example 5-hmC immunofluorescence images of untreated (Con) BMDMs and BMDMs treated with either FAC or desferrioxamine (DFO); scale bars, 100 μm (images are representative of 3 experiments per group). In panels B and C, groups labeled without a common letter were significantly different (P < .05; 1-way ANOVA).