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Blood, Vol. 112, Issue 10, 4268-4275, November 15, 2008
Regulation of the divalent metal ion transporter DMT1 and iron homeostasis by a ubiquitin-dependent mechanism involving Ndfips and WWP2
Blood Foot et al.
112: 4268
Supplemental materials for: Foot et al
Files in this Data Supplement:
- Table S1. siRNA sequences used to knock down Ndfip1 and Ndfip2 (PDF, 166 KB)
- Figure S1. Intracellular localization of DMT1 isoform I (+IRE) (JPG, 124 KB)
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(A) Confocal microscopy shows DMT1-I colocalization with γ-adaptin and both Ndfip1 and Ndfip2. (B) DMTI-I partially colocalises with GM130 and colocalises with Rab5 and Rab11 but not with Rab7. Scale bars represent 10 µm.
- Figure S2. Overexpression of Ndfip1PY or Ndfip2PY reduces transport activity to the same level as that of wild type Ndfip1 or Ndfip2 (JPG, 29.4 KB)
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* indicates a significant difference between transfected and control CHO-DMT1-II cells (p<0.05). Open bar indicates CHO cells not expressing DMT1-II, closed bars indicate CHO-DMT1-II cells. Data expressed as mean ± SEM.
- Figure S3. Immunofluorescence of transfected HA-DMT1-I in untreated CHO cells or in cells treated for 4h with MG132 (50µM) or chloroquine (400µM) (JPG, 36.2 KB)
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Scale bars represent 10 µm.
- Figure S4. Perl’s iron staining of paraffin embedded duodenum sections from wild type and Ndfip1−/− mice (JPG, 44.6 KB)
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Iron (stained black) is seen accumulating in and around the villi in Ndfip1−/− duodenum.
- Figure S5. Morphology of peripheral blood smears from Ndfip1+/+ and Ndfip1−/− mice (JPG, 53.3 KB)
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Mutant mice show no significant difference in red blood cell morphology. Magnification ×40.
- Figure S6. A model of DMT1 trafficking through the cell via its association with Ndfips and WWP2 (JPG, 76.3 KB)
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DMT1 is known to be trafficked through recycling endosomes back to the plasma membrane or internalised and delivered to late endosomes. We propose that at the trans-Golgi network and its associated vesicles (γ-adaptin positive), DMT1 can interact with both Ndfips (dark blue) and WWP2 (pink), which leads to its ubiquitination and promotion of its degradation via the 26S proteasome and/or lysosome.
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