Fig. 5: Tet2OE reverses inhibition of astrogliogenesis by Olig2OE.
From: Tet2-mediated epigenetic drive for astrocyte differentiation from embryonic neural stem cells
a E11 NSCs with Olig2OE and compound expression of Olig2 and Tet2, 6 days after differentiation (Gfap, ctrl vs. Olig2OE, p = 0.022; Olig2OE vs. Olig2OE/Tet2OE, p = 0.014, n = 15 from three independent biological repeats. One-way ANOVA, Tukey test, *P < 0.05, Mean ± SD). Scale bars, 50 μm. b Quantitative PCR analysis of astrocyte markers Gfap and Aqp4 mRNA levels in ctrl NSCs and NSCs with Tet2OE, Tet2KD, Olig2OE, and compound overexpression of Olig2 and Tet2, 5 and 10 days post differentiation. (Gfap, Day5, ctrl vs. Tet2OE, p = 0.00046; ctrl vs. Tet2KD, p = 0.00055; ctrl vs. Olig2OE, p = 0.031; Tet2OE vs. Tet2KD, p = 0.00032; Olig2OE vs. Olig2OE/Tet2OE, p = 0.026; Olig2OE/Tet2OE vs. Tet2OE, p = 0.0066; Olig2OE vs. Tet2OE, p = 0.0006. Day10, ctrl vs. Tet2OE, p = 0.00016; ctrl vs. Tet2KD, p = 0.0088; ctrl vs. Olig2OE, p = 0.0072; Tet2OE vs. Tet2KD, p = 0.00011; Olig2OE vs. Olig2OE/Tet2OE, p = 0.00006; Olig2OE vs. Tet2OE, p = 0.00005, n = 15 from 3 independent biological repeats. Two-way ANOVA, Tukey test, *p < 0.05, **p < 0.01, ***p < 0.001, ns, nonsignificant, Mean ± SD; Aqp4, Day5: ctrl vs. Tet2OE, p = 0.000065; ctrl vs. Tet2KD, p = 0.014; Tet2OE vs. Tet2KD, p = 0.000031; Olig2OE vs. Olig2OE/Tet2OE, p = 0.000294; Olig2OE/Tet2OE vs. Tet2OE, p = 0.029; Olig2OE vs. Tet2OE, p = 0.000632. Day10, Olig2OE vs. Olig2OE/Tet2OE, p = 0.0066, n = 15 from three independent biological repeats. Two-way ANOVA, Tukey test, *p < 0.05, **p < 0.01, ***p < 0.001, ns, nonsignificant, Mean ± SD). c Summary of Tet2 and Olig2 regulation of astrocytic differentiation. DNA demethylation by Tet2 is critical to initiate and establish the transcriptional program that promotes astrocyte differentiation. Olig2 not only represses the expression of astrocytic regulatory factors to block premature astroglial differentiation, it also directly represses Tet2 expression to indirectly maintain DNA hypermethylation astroglial genes.
