Fig. 1

EZH2-deficient hESCs fail to differentiate into NPCs and subtype neuron/glia cells. a Schematic of the default neural differentiation and random differentiation strategy for human embryonic stem cells (hESCs) (more detail information in “Methods” sections). b Morphology of the wild-type (WT) H1 and EZH2^−/− hESCs during neural differentiation at day 0, day 9, day 16 and rosette-like cells. EZH2-deficient hESCs fail to form rosette-like cells. Scale bar, 50 μm. c Immunostaining for the NPC markers PAX6, SOX2, and NESTIN in WT rossettes and EZH2-mutant cells under neural differentiation. Scale bar, 50 μm. d FACS analysis of PAX6⁺ cells at day8 and day16 during neural differentiation in the indicated cells. The data represent the mean ± SD from three independent replicates (n = 3). Significance was determined using unpaired two-tailed Student’s t-tests. **, P < 0.01. e Morphology of the undifferentiated WT NPCs and EZH2-mutant NPCs and their differentiated cells at day 7, day 14 and day 28 during random differentiation. Scale bar, 100 μm. f Immunostaining for the marker of neuron MAP2 and the marker of astrocyte GFAP in the indicted differentiated cells at day 28 from NPCs. Scale bar, 50 μm. Quantity data from MAP2⁺ or GFAP⁺ cells were analyzed. Significance level was determined by unpaired two-tailed Student’s t-tests. ***, P < 0.001. The data represent the mean ± SD (standard deviation) from three independent replicates (n = 3). All error bars throughout the figure represent the SD (standard deviation) from three independent replicates (n = 3)