SEARCH

SEARCH BY CITATION

Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
hep26550-sup-0001-suppfig1.tif4026KFigure S1. Time course analysis of hematocrit and Epo plasma levels after splenectomy in Tg6 mice. Panel A: Tg6 mice overexpressing Epo were splenectomized and sacrificed at different time points. Hematocrit was measured using standard laboratory methods at each time point. Panel B: in a different subset of animals, the concentration of total Epo (e.g. human and mouse) was measured by radio immunoassay in Wt control mice (Wt, n=10), mice overexpressing Epo (Tg6, n=11) and splenectomized Tg6 mice (Tg6_SPL, n=15) two weeks after surgery. Data are presented as mean ± SEM. ** p<0.01 with Wt, ## p<0.01 with Tg6.
hep26550-sup-0002-suppfig2A.tif4617KFigure S2. Flow cytometry and markers of erythropoiesis. Reticulocyte count (Ret), CD71 positive cells and CD44 positive cells were measured in peripheral blood of wild type controls (Wt, n=6), Wt mice treated with 5mg of iron dextran (Wt_DXT, n=5), mice overexpressing Epo (Tg6, n=6), Tg6 mice treated with 5mg of iron dextran (Tg6_DXT, n=5) and Tg6 mice splenectomized (Tg6_SPL, n=4). Panel A: Representative examples of single animals in histograms. Panel B: Quantification of positive cells. Data are presented as mean ± SEM. ** p<0.01 with Wt; # p<0.05 with Tg6; ## p<0.01 with Tg6.
hep26550-sup-0003-suppfig2B.tif733KFigure S2. Flow cytometry and markers of erythropoiesis. Reticulocyte count (Ret), CD71 positive cells and CD44 positive cells were measured in peripheral blood of wild type controls (Wt, n=6), Wt mice treated with 5mg of iron dextran (Wt_DXT, n=5), mice overexpressing Epo (Tg6, n=6), Tg6 mice treated with 5mg of iron dextran (Tg6_DXT, n=5) and Tg6 mice splenectomized (Tg6_SPL, n=4). Panel A: Representative examples of single animals in histograms. Panel B: Quantification of positive cells. Data are presented as mean ± SEM. ** p<0.01 with Wt; # p<0.05 with Tg6; ## p<0.01 with Tg6.
hep26550-sup-0004-suppfig3.tif1524KFigure S3. Liver and spleen iron content. Non-heme iron content was measured in the liver (panel A) and spleen (panel B) of wild type controls (Wt, n=6), Wt mice treated with 5mg of iron dextran (Wt_DXT, n=7), mice overexpressing Epo (Tg6, n=6), Tg6 mice treated with 5mg of iron dextran (Tg6_DXT, n=6) and Tg6 mice splenectomized (Tg6_SPL, n=5). Data are presented as mean ± SEM normalized to Wt controls. ** p<0.01 with Wt, # p<0.05 with Tg6, ## p<0.01 with Tg6.
hep26550-sup-0005-suppfig4.tif2070KFigure S4. Divalent metal transporter 1 expression and intestinal 59Fe absorption. DMT1 expression was measured in the duodenum of wild type controls (Wt, n=6), Wt mice treated with 5mg of iron dextran (Wt_DXT, n=7), mice overexpressing Epo (Tg6, n=6), Tg6 mice treated with 5mg of iron dextran (Tg6_DXT, n=6) and Tg6 mice splenectomized (Tg6_SPL, n=5). Panel A: Western blotting results. Each sample was loaded in 2-4 gels. The panel shows a representative blot obtained with extracts of two animals for each experimental group and the densitometric quantitation of the analysis of the extracts from all of the mice. The DMT1 values were normalized to Wt controls and are presented as mean ± SEM. Panel B: RT-PCR analysis of DMT1 mRNA. Samples from the same animals were analyzed in triplicate and normalized to the housekeeping gene 18S RNA. Data are presented as mean ± 95% CI. Panel C: iron absorption was measured as described in supplemental methods in a different group of Wt (n=4), Wt_DXT (n=5), Tg6 (n=4), Tg6_DXT (n=3) and Tg6_SPL (n=4). Mice were sacrificed 24 hours post-gavage and the gastrointestinal tract was removed and radioactivity was measured using a gamma counter. Percent 59Fe absorption was calculated, normalized to Wt controls and presented as mean ± SEM. * p<0.05 with Wt; ** p<0.01 with Wt; # p<0.05 with Tg6.
hep26550-sup-0006-suppfig5.tif1001KFigure S5. Analysis of Ferroportin mRNAs. Ferroportin IRE (Panel A) and non-IRE (Panel B) transcripts were measured in wild type controls (Wt, n=6), Wt mice treated with iron dextran (Wt_DXT, n=7), mice overexpressing Epo (Tg6, n=6), Tg6 mice treated with iron dextran (Tg6_DXT, n=6) and splenectomized Tg6 mice (Tg6_SPL, n=5) by RT-PCR. Samples were analyzed in triplicate and normalized to the housekeeping gene 18S RNA. Data are presented as mean ± 95% CI. ** p<0.01 with Wt; p=0.055 with Wt.
hep26550-sup-0007-suppinfo.doc94KSupporting Information

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.