Fig. 5. CHD8 loss-of-function disrupts neuroectoderm differentiation.

A Cartoon schematic showing the protocol of differentiating human ESCs toward neural fate. B the dynamic gene expression of CHD8 during neuroectoderm differentiation (n = 3). C RT-qPCR analysis of neuroectoderm genes PAX6 and SOX1 and pluripotent gene NANOG at day 0, 3, 5, 7 of differentiation (n = 3). Wildtype ESCs at day 0 served as control. D, E Immunofluorescence of neuroectoderm markers SOX1 and PAX6 in wildtype and knockout groups at day 3 (D) and day 7 (E) of differentiation. The percentage of SOX1- or PAX6-positive cells was calculated (n = 4). Scale bar = 200 µm. F GO enrichment analysis for downregulated genes in CHD8 knockout NPC (P < 0.05, log₂(fold-change) < −1). The error bars represented the mean ± SD and the significance level was calculated by Student’s t-test (two-tailed, equal variance) (ns means not statistically significant, *P < 0.05, **P < 0.01, ***P < 0.001).