Characterizing in vitro neural stem cells to study Methyl CpG binding protein 2 (MeCP2) expression. (A) Schematic representation of in vitro neural stem cell (NSC) differentiation. (B) Detection of (a) NESTIN+ and (b) KI67+ cells in self-renewing neurospheres. Scale bars represent 20 μm. (C) Immunofluorescent detection of different cell-type markers in the day 8 (D8) population (a) TUBULIN III (TUB III): neurons, (b) Glial fibrillary acidic protein (GFAP): astrocytes, (c) S100B: mature astrocytes, (d) 2',3'-Cyclic-nucleotide 3'-phosphodiesterase (CNPase): oligodendrocytes, (e) Myelin basic protein (MBP): oligodendrocytes, (f) Oligodendrocyte lineage transcription factor 2 (OLIG2): early oligodendrocytes and progenitors, and (g) KI67: proliferating cells. Scale bars represent 10 μm. The percentages represent average number of cells from three individual experiments (n = 3 ± standard error of the mean). (D) (a) Immunofluorescent detection of MeCP2 in a sectioned primary neurosphere. Scale bar represents 20 μm. (b) Double labeling of MeCP2 and NESTIN within primary neurosphere cells. Scale bar represents 5 μm. (E) Immunofluorescent detection of MeCP2 in D8 cell types: (a) TUB III, (b) GFAP, (c) S100B, (d) CNPase, (e) MBP, (f) OLIG2, and (g) KI67. Scale bars represent 2 μm.