SUPPLEMENTARY FIG. S2. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of dnscs.
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1 Supplementary Data SUPPLEMENTARY FIG. S1. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of anpcs. A panel of lineage-specific markers were used to quantify the pattern of neural differentiation in cultures of anpcs after 1 week in 1% FBS. The markers used were GFAP for the astrocyte lineage, Olig2 and PDGFRa for the oligodendrocyte lineage, biii tubulin for the neuronal lineage, and nestin expression was used to identify undifferentiated anpcs. anpc, adult neural precursor cell; FBS, fetal bovine serum; GFAP, glial fibrillary acidic protein; PDGFRa, platelet-derived growth factor alpha; DAPI, 4,6-diamidino-2-phenylindole; YFP, yellow fluorescent protein; Olig2, oligodendrocyte transcription factor 2.
2 SUPPLEMENTARY FIG. S2. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of dnscs. A panel of lineage-specific markers were used to quantify the pattern of neural differentiation in cultures of dnscs after 1 week in 1% FBS. The markers used were GFAP for the astrocyte lineage, Olig2 and PDGFRa for the oligodendrocyte lineage, biii tubulin for the neuronal lineage, and nestin expression was used to identify undifferentiated dnscs. Scale bar represents 80 mm.
3 SUPPLEMENTARY FIG. S3. Histological confirmation of a lack of compact myelin and MBP expression in the shiverer mouse CNS. A mutation resulting in the loss of expression of MBP results in the loss of compact CNS myelin. (A) H&E and luxol fast blue (LFB) staining for observation of compact myelin in spinal cord cross sections and (B) longitudinal sections of dorsal column white matter were assessed using fluorescent immunohistochemistry for MBP (green) and NF200 (red) and show the association of MBP and axons in the spinal cord white matter of a wild-type mouse and the loss of MBP in the shiverer mutant. Scale bar represents 15 mm. MBP, myelin basic protein; CNS, central nervous system; H&E, hematoxylin and eosin.
4 SUPPLEMENTARY FIG. S4. Sample counting fields for oligodendrocyte lineage markers used in quantification of the in vivo neural differentiation of pattern of anpcs 6 weeks after transplantation into the spinal cord of shiverer mice. Typical image fields used for cell quantification are shown for transplanted anpcs stained for the oligodendrocyte lineage markers Olig2, APC, and PDGFRa. Arrowheads indicate cells in which the lineage marker is observed to colocalize with YFP of the transplanted anpcs. Scale bar represents 15 mm. APC, adenomatous polyposis coli.
5 SUPPLEMENTARY FIG. S5. Sample counting fields for astrocyte, neuronal, and neural stem cell lineages used in quantification of the in vitro neural differentiation of pattern of anpcs 6 weeks after transplantation into the spinal cord of shiverer mice. Typical image fields used for cell quantification are shown for transplanted anpcs stained for the astrocyte lineage marker GFAP, the neuronal lineage marker MAP2, and nestin. Arrowheads indicate cells in which the lineage marker is observed to colocalize with YFP of the transplanted anpcs. Scale bar represents 15 mm. MAP2, microtubule-associated protein 2.
6 SUPPLEMENTARY FIG. S6. Sample counting fields for astrocyte, neuronal, and neural stem cell lineages used in quantification of the in vivo neural differentiation of pattern of dnscs 6 weeks after transplantation into the spinal cord of shiverer mice. Typical image fields used for cell quantification are shown for transplanted dnscs stained for the astrocyte lineage marker GFAP, the neuronal lineage marker MAP2, and nestin. Arrowheads indicate cells in which the lineage marker is observed to colocalize with YFP of the transplanted dnscs. Scale bar represents 15 mm.
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