International Journal of Stem Cells : eISSN 2005-5447

Fig. 4.

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Fig. 4. Shh-independent activation by SAG restored motor neuron differentiation in Cdo-deficient ESCs. To activate Shh signaling pathway in Shh-independent way, Cdo+/+ and Cdo−/− ESCs were treated with 1 μM SAG instead of Shh during MNS stage. (a, b) qRT-PCR analysis for genes related to ventral neural subtypes and motor neuron specification (Pax6, Olig2, Nkx6.1, Hb9, Isl1 and Chx10) (a) and dorsal interneuron specification (Brn3a, Pax2, Lbx1 and Pax3) (b) in Cdo+/+ and Cdo−/− ESCs at MNS-5 and Elong. L32 was selected as a reference gene. Data represent means±SEM. *p<0.05, **p<0.01, ***p<0.001. (n=3, each) (c) Immunostaining for Olig2/Nkx6.1or Hb9/Isl1 in Cdo+/+ and Cdo−/− EBs at MNS-5. DAPI was used to visualize nuclei. Size bar=25μm. (d) Quantification of Olig2-, Nkx6.1-, Hb9- and Isl1-positive cells shown in panel c. Data represent means±SD. *p<0.05. (n=6, each) (e) Immunostaining for Hb9 or Isl1 in Cdo+/+ and Cdo−/− cells at Elong. NF was immunostained indicating neuronal differentiation. Nuclei were visualized by DAPI. Size bar=10μm. (f) Quantification of Hb9- or Isl1-positive cells shown in panel e. Data represent means±SD. *p<0.05 (n=6).
International Journal of Stem Cells 2020;13:342-52
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