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Blood, Vol. 112, Issue 4, 1503-1509, August 15, 2008
Iron regulates phosphorylation of Smad1/5/8 and gene expression of Bmp6, Smad7, Id1, and Atoh8 in the mouse liver
Blood Kautz et al.
112: 1503
Supplemental materials for: Kautz et al
Files in this Data Supplement:
- Table S1. Sequences of the primers used for validation of microarray results by real-time quantitative PCR (PDF, 49.3 KB)
- Table S2. Functional categories over-represented in groups of genes differently regulated by iron overload and/or iron deficiency in the liver (PDF, 27.5 KB) -
Genes in the different groups were compared with the whole set of mouse genes for over-representation of functional categories, using the DAVID (Database for Annotation, Visualization, and Integrated Discovery) annotation tool. The Category column shows the original database/resource from which the terms originate. The Term column indicates enriched terms associated with the gene list. The n column indicates the number of genes involved in the term. The expression analysis systematic explorer (EASE) score is a modified Fisher exact P value. BP, biological process; CC, cellular component; MF, molecular function; GO, Gene Ontology. There was no significant over-represented functional category among the 154 and 105 transcripts whose expression was found either down-regulated or up-regulated in both conditions, respectively.
- Table S3. Genes included in the functional categories listed in Table S2 (PDF, 78 KB)
- Figure S1. Expression of Bmp2 and Bmp4 in the liver of mice fed diets with different iron contents (JPG, 20.7 KB)
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The expression of Bmp2 and Bmp4 transcripts was assessed in mice of two strains (B6 and D2) fed an iron-deficient (ID), iron-balanced (IB) or iron-enriched (IE) diet. Expression levels were normalized to HPRT and log2 transformed, and are expressed as means of five samples ± standard deviation. The contribution of diet was assessed by ANOVA and statistical significant differences relative to the iron-balanced diet were determined by Sheffe post-hoc tests. ***p < 0.001.
- Figure S2. Dietary iron content modulates liver Id1 protein levels (JPG, 41.9 KB)
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Liver lysates from mice fed an iron-deficient (ID), iron-balanced (IB) or iron-enriched (IE) diet (N=3 in each group) were analyzed by western blot with an antibody to Id1. Blots were stripped and reprobed with an antibody to β-actin as loading control. A representative experiment is shown.
- Figure S3. Smad1/5/8 phosphorylation relative to Smad1 expression (JPG, 83.9 KB)
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Liver lysates from mice fed an iron-deficient (ID), iron-balanced (IB) or iron-enriched (IE) diet (N=3 in each group) were analyzed by western blot with an antibody to phosphorylated Smad1/5/8. Blots were stripped and reprobed with an antibody to Smad1 as loading control. A representative experiment is shown. Although Smad1 expression is much lower in D2 mice than in B6 mice, within each strain the ratio of phosphorylated Smad1/5/8 relative to Smad1 is increased by iron overload and decreased by iron deficiency.
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