Fig. 1: Tet2 positively regulates the expression of astroglial lineage genes.

a Spontaneous differentiation of astrocytes from Dnmt3a-/- embryonic NSCs in the presence of bFGF. Scale bars, 100 μm. b DNA demethylation of the GFAP proximal promoter in Dnmt3a-/- NSCs. c Tet 1, 2, 3 mRNA expression in undifferentiated NSCs, fully differentiated astrocytes, and immature neurons isolated from E15 mouse cortices (Tet1, NSC vs astrocyte, p = 0.030; NSC vs neuron, p = 0.032; Tet2, NSC vs neuron, p = 0.044; astrocyte vs neuron, p = 0.039; Tet3, astrocyte vs. neuron, p = 0.037; n = 12, 3 independent biological repeats. One-way ANOVA, LSD test, *p < 0.05, ns, nonsignificant, Mean ± SD). d Tet2 mRNA levels during NSC spontaneous differentiation after bFGF withdrawal at Day 0, 2, 4, and 6. (Day2 vs Day0, p = 0.028; Day6 vs Day0, p = 0.0044; n = 12, 3 independent biological repeats. One-way ANOVA, Tukey test, *p < 0.05, **p < 0.01, ns, nonsignificant, Mean ± SD). e Exogenous Tet2 catalytic domain overexpression (Tet2OE) induced demethylation of Gfap promoter. Methylation levels at two proximal promoter segments near the TSS (1, 2) and distal promoter segment (3) in undifferentiated NSCs with control and Tet2OE conditions (ctrl vs. Tet2OE (1) p = 0.0044; (2) p = 0.021; n = 15, from three independent biological experiments (Two-tailed Student’s t-test, *p < 0.05, **p < 0.01, ns, nonsignificant, Mean ± SD). f Top: A Pairwise comparisons with Pearson correlation coefficients of transcriptomes from control and Tet2OE before and after differentiation at day 3. Red indicates a high similarity (cor = 1) and white indicates a lower similarity (cor = 0) between the two samples. Bottom: CummeRbund volcano plot reveals genes that are significantly differentially expressed between each pair. g Top 25 upregulated genes by Tet2OE in NSCs at day 3 following differentiation. Green indicates astrocyte lineage genes; blue indicates oligodendrocyte lineage genes and red indicates neuronal genes.