Fig. 8

Enhanced KIF2A expression partially rescued deficits in cilium length, NPC proliferation, and organoid sizes. a, c Confocal imaging of CEP170 knockdown (a) or WDR62 mutant (c) human NPCs stained with antibodies against Arl13b (red) and KIF2A (green). Hoechst stains nuclei (blue). Scale bars: 2.5 μm. b, d Quantification of KIF2A signal intensity. e Confocal imaging of control or KIF2A-GFP lentivirus-infected human NPCs stained with antibodies against Arl13b (red) and KIF2A (green). Hoechst stains nuclei (blue). Scale bars: 1 μm. f Quantification of cilium length in e. g Confocal imaging of BrdU-positive human NPCs (red). Hoechst stains nuclei (blue). Scale bars: 25 μm. h–j Quantification of the percentage of BrdU, Ki67, and Β3-Tubulin-positive cells out of total human NPCs. k Quantification of surface areas of cerebral organoids. l Quantification of p-H3-positive cells in VZ-like regions of mutant organoids with or without KIF2A overexpression. For all the experiments, error bars represent SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, n.s. represents not significant (Student’s t-test). m Diagram showing that a WDR62-CEP170-KIF2A microcephaly protein pathway promotes cilium disassembly, disruption of which reduces NPCs and contributes to microcephaly