Sox6 overexpression decreased the multi-lineage differentiation potential in NSPCs.

<p>(A) Neurospheres infected with either a control retrovirus expressing GFP or a retrovirus expressing Sox6 and GFP were cultured for 5 days in the presence of EGF and FGF2. The neurospheres were allowed to dissociate and were then plated onto poly-L-ornithin-coated coverslips at a density of 2×10⁵ cells/cm² and cultured for another 5 days without growth factors. Fully differentiated cells were fixed and subjected to immunocytochemical analyses. The number of GFP-positive cells that were co-labeled with a neuronal marker (TuJ1), an astrocyte marker (GFAP), or an oligodendrocyte marker (CNPase) was counted and the data were expressed as percentages of GFP-positive cells. The graph represents the average of two independent experiments. (B) Representative images of cells that differentiated from neurospheres infected with a control retrovirus expressing GFP (Ctrl) or a retrovirus expressing Sox6 and GFP (Sox6). Scale bar: 20 µm. (C, D) Retroviral Sox6 overexpression in NSPCs led to an increase in Nestin (C) and Musashi-1 (D) gene expression 5 days after infection. Data are representative of three independent experiments. Error bars indicate S.D. values; **<i>P</i><0.01 versus control; Student’s <i>t</i>-test.