Figure 5

Impairment at the early stage of neural differentiation of TET1-deficient hESCs under the morphogen-free condition. (a) Protocol for the differentiation of embryoid bodies (EB) to early neural lineage under morphogen-free condition from day 0 to day 14. N2, N2 supplements. (b–d”) Three types of neuroepithelial colonies were classified by the percentage of SOX1⁺ NESTIN⁺ cells among all cells within individual colony, representing the completeness of neural lineage commitment. Full, colonies with a SOX1⁺NESTIN⁺ percentage >70% (b–d for separate channels and merged image); Partial, colonies with a SOX1⁺NESTIN⁺ percentage between 20–70% (b’-d’); Limited, colonies with a SOX1⁺NESTIN⁺ percentage <20% (b”-d”). Bar, 500 µm. (e) Percentage of each type of colonies formed at D14 by WT and the two TET1-deficient lines of H9 hESCs. *p < 0.001; Fisher’s exact test, n = 40 colonies for each cell line.