Stabilization of induced neural stem cells with tet-On transactivator gene.
Representative images of (A) undifferentiated iNSc tet expressing neural precursor markers SOX2 (green) and nestin (red), and (B) differentiated iNSc tet expressing astroglia marker GFAP (green) and neuronal-specific marker Map2 (red). Cell nuclei were counterstained with the nuclear DNA marker DAPI (blue). Scale bars represent 50 μm (A-B). (C) Induction of green fluorescent protein (GFP, green) expression in iNSc tet-GFP treated with doxycycline (1 μg/mL, Dox) for 96 hours. Cell nuclei were counterstained with the nuclear DNA marker DAPI (blue). Scale bars represent 100 μm. (D, E) Quantification of GFP fluorescence intensity in iNSc tet-GFP after 48 and 72 hours of incubation with doxycycline at different concentrations. Statistical significance calculated by One-way ANOVA with Dunnett’s multiple comparison test. Error bars indicate SEM. **, p<0.01; ***, p<0.005; ns, not significant.