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. 2016 Aug 26;23(12):2042–2053. doi: 10.1038/cdd.2016.85

Figure 4.

Figure 4

Figure 4

Knockdown of Stox1 enhances proliferation and delays the differentiation of GNPs. (a) Quantitative RT-PCR and immunobloting confirm efficient silencing of Stox1 in shStox1 adenovirus-infected GNPs. (b) Infection of GNPs isolated from Dcx-DsRed mice with shStox1 adenovirus leads to decreased generation of Dcx-DsRed- positive cells. Representative flow cytometric plots are shown. Histograms show the percentage of Dcx-dsRed-positive cells. (n=3). (c). Immunofluorescent staining indicates that Stox1 knockdown represses the differentiation of NeuN-positive granule neurons from in vitro culture of GNPs. The scale bar represents 25 μm. (d) Stox1 knockdown downregulates the differentiation of Dcx-positive granule neurons from in vitro culture of GNPs. The scale bar represents 25 μm. (e) The mRNA levels of Math1 are upregulated, while Dcx, Tuj1, NeuN, Neurod1, Zic1 and Zic2 are downregulated in shStox1 adenovirus-infected GNPs. (f) Knockdown of Stox1 in GNPs leads to increased cell proliferation in the presence of SHH (1μg/ml). Cell proliferation is assessed by EdU incorporation assay. (n=3). (g) Quantitative RT-PCR analysis demonstrates enhanced expression of Ccnd1 and reduced mRNA levels of p18, p21 and p27 in shStox1 infected GNPs cultured in the presence of 1μg/ml SHH. (***P<0.001, **P<0.01,*P<0.05, data are shown as means±S.E.M.)