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. 2016 Jan 26;26(2):171–189. doi: 10.1038/cr.2016.15

Figure 5.

Figure 5

SOX2 KD leads to impaired neuronal differentiation due to the increased activity of canonical Wnt signaling and reduced expression of pro-neuronal genes. (A) Western blot for p-LRP6, LRP6 and active caspase3 on day 3 of neuronal differentiation from hNPCs. SOX2 KD was conducted on day 1. ChIR99021 (3 μM) was used as a positive control. (B) RT-qPCR results of WNT pathway genes (left panel), and neuronal markers (right panel) on day 3 of SOX2 KD under neuronal differentiation conditions in the presence of DMSO, or IWR-1e (10 μM), or ChIR99021 (3 μM). The values in the control (siNT+DMSO) are set as 1. Statistical analyses are made by comparing control and experiment groups. (C) Immunostaining of MAP2, TUJ1 and DCX upon ChIR99021 treatment or SOX2 KD under neuronal differentiation conditions. SOX2 KD was carried out in the beginning of neuronal differentiation (day 1). IWR1-e and ChIR99021 were added from day 2 to day 7. Immunostaining was performed on day 7. (D) RT-qPCR results of pro-neuronal genes (NEUROD1 and NEUROG1) under the conditions in B. (E) ChIP-qPCR results of SOX2 enrichments at promoters of NEUROD1 and NEUROG1 in hESCs and hNPCs.