Generation of a human haploid neural stem cell line for genome-wide genetic screening

Figure 3 Identification and differentiation of haploid neural stem cells. A: Immunofluorescence analysis of the neural stem cell (NSC)-specific markers Nestin (red), PAX6 (green), and SOX1 (red) in derived haploid NSCs (haNSCs). DAPI (blue) was used to stain the nuclei. Scale bar, 50 μm; B: Immunofluorescence analysis showed that haploid neural spheres expressed NSC-specific proteins Nestin (red), PAX6 (green) and SOX1 (red). Scale bar, 100 μm; C: Transcript levels of diploid NSCs, monolayer cultured haNSCs, haploid neural spheres, human haploid embryonic stem cells (hPGES1) and human fibroblasts; D: Real time PCR analysis of NSC marker genes (SOX1, PAX6, and Nestin); E: Fluorescence activated cell sorting of derived astrocytes, oligodendrocytes, and neurons by DNA content; F: Immunofluorescence analysis of astrocytes (GFAP, green) and oligodendroglia (O4, red). DAPI (blue) was used to stain the nuclei. Scale bar, 50 μm; G: Immunofluorescence analysis of Tuj1 (green) and Map2 (red) in neurons derived from haNSCs. DAPI (blue) was used to stain the nuclei. Scale bar, 50 μm. ^bP < 0.01; ^cP < 0.001.