Fig. 3: KDM6s-deficient NPCs fail to differentiate into neurons and glia.

a Strategic diagram of the spontaneous differentiation of human NPCs. Wild-type (WT) NPCs or three KDM6 mutant NPC lines lacking UTX, JMJD3 or both at passage 2 (NPC-P2) were plated in differentiation media lacking FGF2 and EGF and cultured for 28 days for differentiation. b Morphology of the undifferentiated WT NPCs or three KDM6 mutant NPC lines and their differentiated cells at day 28. Scale bar, 50 μm. c Immunostaining for the neuronal marker MAP2 and the glial marker GFAP in the indicted undifferentiated or differentiated NPCs. Scale bar, 50 μm. Quantity data from MAP2⁺ or GFAP⁺ cells were analysed. Significance level was determined by unpaired two-tailed Student’s t-tests. **P < 0.01. The data represent the mean ± SD (standard deviation) from three independent replicates (n = 3). d Immunostaining for the NPC markers SOX2/NES in the indicated undifferentiated or differentiated NPCs. Scale bar, 50 μm. The percentage of SOX2⁺ or NES⁺ cells was analysed. Significance was determined by unpaired two-tailed Student’s t-tests. **P < 0.01. The data represent the mean ± SD from three independent replicates (n = 3). e qRT-PCR analysis of the expression of the NPC markers NES/SOX2/SOX1/PAX6, the neuronal markers DLX2/DLX1/MAP2/TUBB3/NEUN/SCL31A1 /GBJ6, the glial marker GFAP, and UTX/JMJD3 in the indicated differentiated cells at day 28. The significance level was determined using unpaired two-tailed Student’s t-tests. **P < 0.01. The data represent the mean ± SD from three independent replicates (n = 3). f Spearman’s rank correlation analysis of the whole-genome transcriptome of the indicated undifferentiated or differentiated NPCs. All error bars throughout the figure represent the SD (standard deviation) from three independent replicates (n = 3). Source data are provided as a Source Data file. See Supplementary Fig. 3 for more information.