The Journal of Neuroscience, January 25, 2006, ():
A Critical Role of Erythropoietin Receptor in Neurogenesis and Post-Stroke Recovery
J. Neurosci. Tsai et al.
26: 1269
Supplemental data
Files in this Data Supplement:
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Supplemental material
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Supplementary Figure 1. Epo and EpoR are necessary for embryonic neurogenesis. Whole-mount in situ hybridization with EpoR (a, c, e) or Epo (b, d, f) anti-sense probes. EpoR is expressed throughout the neural fold by E8.5, except where the neural fold has begun to fuse (Supp Fig. 1a, arrow). By E9.5, EpoR expression is enhanced in the forebrain and optic placode (op, Supp Fig. 1c). Epo is expressed in similar regions to EpoR, but its expression trails EpoR’s by 0.5-1 day: Epo expression is absent at E8.5 (Supp Fig.1b) but is found in the op and in the mesencephalon at E9.5 (Supp Fig.1d, arrows). At E10.5, Epo expression spreads in the mesocoel region and is enhanced in the optic vesicle (Supp Fig.1f) while EpoR expression is apparent in the mandible arch and the hyeloid arch, along the neural crest migration pathway (Supp Fig. 1f). Aberrant neurogenesis in EpoR null animals at E10.5 (g,h) and E13.5 (I-l) E13.5. Red arrows denote open neural tube at E10.5, and wavy, underdeveloped neural tube at E13.5. Arrows indicate the under-developed telencephalic region in the EpoR null animal.
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Supplementary Figure 2 Targeting and generation of floxed EpoR allele. (a) Schema for generation of floxed EpoR allele. Southern probes shown. (b) Genomic DNA was extracted from ES clones and digested by EcoRV. Southern blots were probed with either 5' probe (left panel) or 3' probe (right panel) as indicated in A.
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Supplementary Figure 3 Doublecortin positive cells in SVZ, subcortical white matter and cortex in control and after stroke. (a) Control brain. Doublecortin positive cells are present in the SVZ and as clusters just lateral to the SVZ and closely adjacent white matter, (b) Three days post-stroke. Many doublecortin positive cells extend between the SVZ and cortex, filling the subcortical white matter. (c) Three days post-stroke at higher magnification. In a different case from panel b, doublecortin positive cells can be seen streaming from the SVZ into the white matter. These cells have a bipolar, migratory appearance with elongated nuclei and linear horizontal processes. (d) Seven days post-stroke. Doublecortin positive cells are present in cortex and exhibit a different morphology, with more rounded nuclei and irregular local processes. Bar in b,d = 25uM.
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Supplementary Figure 4 BrdU and doublecortin double fluorescent immunohistochemistry. Sections are taken from peri-infarct cortex at days 5 and 7 post-stroke. Arrows indicate double-labeled cells in the merged images. Bar = 30uM.
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Supplementary Figure 5 Migrating cells after stroke display markers of immature neurons and progenitor cells. Images in top two rows are taken from peri-infarct cortex. Images from bottom two rows are taken from subcortical whit matter. Bar = 30uM.
