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

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SC_12-0172_sm_supplFigure1.pdf345KSupplementary Fig. 1. Floor plate progenitors induced by SHH or CHIR. (A) The floor plate progenitors exhibited epithelial morphology in each individual colony on day 8. (B) Immunostaining for FoxA2, Otx2, and En1 at day 18 after treatment with 500ng/ml SHH. (C) Immunostaining for En1 (green), HoxA2 (red) and HoxB1 (blue) at day 18 after treatment with 2 or 3 uM of CHIR. (D) Immunostaining for En1 (green), OTX2 (red) and FOXA2 (blue) at day 18 after treatment with 1 uM of CHIR. (E) Immunostaining for TH (green) and 5HT (red) at day 36 after treatment with 0.4 or 1 uM of CHIR and subsequent treatment with FGF8. (F) Quantification of the data presented in E. (G) Immunostaining for TH (green) and GABA (red) at day 36. (H) Immunostaining for TH (green) and NESTIN (red) at day 36. Bar =50um
SC_12-0172_sm_supplFigure2.pdf491KSupplementary Fig. 2. Midbrain floor plate progenitors induced by SHH and CHIR. (A) Separate immunofluorescent channels correspond to merged images in Fig. 3d. (B) Immunostaining of midbrain floor plate progenitors for Nestin (green), Lmx1a (red) and FoxA2 (blue) at day 18. (C) Immunostaining for Ngn2 (green) and Mash1 (red) at day 22 after withdrawal of SHH and CHIR from day 13. Bar =50um
SC_12-0172_sm_supplFigure3.pdf311KSupplementary Fig. 3. Differentiation of midbrain floor plate progenitors and DA neurons from human iPSCs (line IMR90-4). (A) Immunostaining and quantification of markers for midbrain floor plate progenitors at day 18 (Corin, FoxA2, En1, Lmx1a and Otx2). (B) Co-staining of TH (green) with other midbrain DA neuronal markers FoxA2, En1, Lmx1a and Nurr1 (red) as well as quantification of the cell populations. Bar =50um
SC_12-0172_sm_supplFigure4.pdf230KSupplementary Fig. 4. Electrophysiological properties of DA neurons. (A) Electrophysiological characteristics of neurons that were differentiated for 10 weeks in vitro. (B) Inward Na+ and outward K+ currents were triggered upon - 50mV to + 50 mV voltage steps. The initial currents were enlarged in the lowest panel. (C) Action potentials were induced from − 60 pA to + 60 pA injected current steps. The lower panel shows the response upon + 60 pA current injection. (D) Both excitatory (inward current) and inhibitory (outward current) spontaneous postsynaptic currents are present which are inhibited by CNQX and bicuculine, respectively. The asterisks indicate individual excitatory (sEPSCs) and inhibitory (sIPSCs) events.
SC_12-0172_sm_supplFigure5.pdf800KSupplementary Fig. 5. Differentiation of midbrain floor plate progenitors and DA neurons from rhesus monkey iPSCs. (A) Experimental paradigm showing differentiation of midbrain DA neurons from rhesus monkey iPSCs. (B) Immunostaining of markers for midbrain floor plate progenitors at day 18 (Corin, FoxA2, En1, Lmx1a and Otx2). (C) Co-staining of TH (green) with other midbrain DA neuronal markers FoxA2, En1, Lmx1a and Nurr1 (red). (D) Quantification of midbrain floor plate progenitor populations among total cells. (E) Quantification of TH+ DA neuron populations and midbrain marker expression in TH+ cells. Bar =50um
SC_12-0172_sm_supplTable1.pdf286KSupplementary Table

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