For imaging, all pictures were taken using a Leitz microscope interfaced with a Spot RT color camera. Fluorescent intensity observations were determined independently by consensus of two analyzers based on the following criteria. To verify the intra-cellular and intra-nuclear distribution of the epigenetic marks, we have examined the cells under both fluorescent- and phase-contrast bright-field microscopy, alternatively or simultaneously (with dimmed bright-field). Positive stained neurospheres were stained with 5-MeC (Normal, N=12; Alcohol, N=10) or DMNT1 (Normal, N=12; Alcohol N=10), with fluorescent intensity compared to the dark background. Based on the rank values, Mann-Whitney U was used for nonparametric statistical analysis (StatView, Cary, NC) of ranked fluorescent intensity to comparing each neurosphere compartment to the undifferentiated neurospheres.
