RNeasy Mini Kit (Qiagen, Ontario, Toronto, Canada) was used for RNA extraction as per the manufacturer’s protocol. Preparation of cDNA and qRT-PCR were carried out as described previously [40-42]. Transcript levels of Mecp2 (total), Mecp2e1 [NCBI: NM_001081979.1], Mecp2e2 [NCBI: NM_010788.3], Dnmt genes (Dnmt1, Dnmt3a and Dnmt3b), neuronal genes (Tubulin III (Tub III), NeuN), astrocytic genes (Gfap, S100b), and oligodendrocyte-specific genes (Cnpase, Mbp) were examined by using gene-specific primers (Table 2), as described previously [37,43]. The relative expression and fold changes were calculated as described previously [37]. Two-way analysis of variance (ANOVA) and the Student t-test were used to calculate significant differences between untreated control and decitabine-treated samples.
