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

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STEM_1049_sm_SuppFig1.pdf102KSupplementary Figure 1. Temporal pattern of expression of sFRP1-4 in the developing ventral and dorsal midbrain. (A-D) Quantitative RT PCR analysis was performed with rat ventral midbrain (vm) tissue obtained at embryonic days 10.5, 11.5, 13.5 and 15.5, and compared to dorsal midbrain (dm). Note that the initially high levels of sFRP1-3 mRNAs (A-C) are downregulated by the end of the neurogenic period. SFRP4 mRNA levels remained low and were not temporally regulated (D). *** p<0.001, ** p<0.01 and * p<0.05 compared to vm or dm at E10.5, respectively. One-way ANOVA, with Bonferroni's post-hoc test for multiple comparisons (N=3).
STEM_1049_sm_SuppFig2.tif754KSupplementary Figure 2. High concentrations of sFRP2 have no effect on basal or Wnt5ainduced DA differentiation of VM primary precursor cultures. (A) Rat E14.5 ventral midbrain cultures were treated for 3 days with vehicle (BSA Ctrl), sFRP1 or sFRP2 (5μg/ml) and analyzed for expression of beta III-tubulin, (TuJ1 - red) and tyrosine hydroxylase (TH - green) and counterstained with Hoechst (blue). (B) The number of TuJ1+ and TH+ neurons decreased in sFRP1-treated cultures. (C) Neither Wnt5a nor SFRP2 had any effect on the number of neurons. (D) While Wnt5a increased the number of TH+ cells, sFRP2 had no effect on the number of TH+ cells and did not modulate the effects of Wnt5a. * p<0.05 One-way ANOVA, with Bonferroni's post-hoc test for multiple comparisons (N=3).
STEM_1049_sm_SuppFig3.tif1302KSupplementary Figure 3. Proliferation in sFRP1−/−sFRP2−/− mice. (A) The number of A9-A10 VM TH+ cells was not altered in sFRP1−/−;sFRP2−/− embryos, compared to wild-type embryos at E14.5. (B) Quantification of Ki67 revealed no difference in the number of Ki67+ cells in WT and sFRP1−/−sFRP2−/− VM. (C) Stainings for Ki67 and Tuj1 of showed no detectable difference between sFRP1−/− and littermates sFRP1−/−;sFRP2−/− mice in the total number of neurons or cells in the cycle.
STEM_1049_sm_SuppInfo.pdf90KSupplementary Data

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