Fig 2: ASD-related de novo mutations in POGZ impair the nuclear localization of the POGZ protein.a De novo mutations in POGZ identified in patients with NDDs (blue, ASD; green, ID; red, both ASD and ID; purple, unclassified NDDs). b De novo mutations in POGZ identified in unaffected controls. c The number of missense, nonsense, and frameshift mutations identified in patients with NDDs and unaffected controls. d Distribution of POGZ mutations identified in patients with NDDs and unaffected controls (blue, missense mutations; red, nonsense mutations; green, frameshift mutations). e, g ASD-related nonsense e or missense g mutations disrupted the nuclear localization of Myc-tagged overexpressed mouse (m) POGZ in Neuro2a cells. C cytosolic fraction; N nuclear fraction; WT wild-type. Note that Q1038R mutation in mouse POGZ corresponds to Q1042R mutation in human POGZ. f, h, j Quantification of POGZ in cytosolic and nuclear fractions (each n = 4). i Control mutations identified in unaffected healthy controls did not affect the nuclear localization of Myc-tagged overexpressed mouse POGZ in Neuro2a cells. C cytosolic fraction; N nuclear fraction; WT wild-type. Note that the amino acid numbers are based on the human protein (NP_055915.2) a, b and mouse protein (NP_766271.2) e, g, i. f, h, j One-way ANOVA with Bonferroni–Dunn post hoc tests, f F3, 12 = 23.12; h F2, 9 = 67.45; j F3, 12 = 0.491. ***P < 0.001. Data are presented as the mean ± s.e.m. The source data underlying figures e, g and i are provided as a Source Data file.

Fig 3: POGZ regulates the neuronal differentiation of mouse cortical neural stem cells.a Slight, non-significant migration defects caused by shRNA-mediated knockdown of Pogz in E16.5 mouse cortices electroporated at E14.5. CP cortical plate; IZ intermediate zone; SVZ subventricular zone; VZ ventricular zone; E embryonic day. Scale bars, 50 µm. b Quantification of GFP+ cells in each layer (each n = 4). CP cortical plate; IZ intermediate zone; SVZ subventricular zone; VZ ventricular zone. c Increased number of PAX6+ NSCs caused by Pogz knockdown in E16.5 mouse cortices electroporated at E14.5. d Quantification of PAX6+ cells (each n = 4). e Decreased number of TBR2+ differentiated IPs caused by Pogz knockdown in E16.5 mouse cortices electroporated at E14.5. f Quantification of TBR2+ cells (each n = 4). g Decreased number of SATB2+ differentiated neurons caused by Pogz knockdown in E16.5 mouse cortices electroporated at E14.5. h Quantification of SATB2+ cells (each n = 4). c, e, g Right panels, magnifications of the areas outlined with orange boxes. Arrowheads indicate co-labeled cells. White scale bars, 50 µm; orange scale bars, 10 µm. b Two-way repeated-measures ANOVA with Bonferroni–Dunn post hoc tests, F4, 27 = 1.861. d, f, h One-way ANOVA with Bonferroni–Dunn post hoc tests; d F2, 9 = 18.37; f F2, 9 = 5.710; h F2, 9 = 11.91. *P < 0.05, **P < 0.01. Data are presented as the mean ± s.e.m.

Fig 4: POGZ regulates the mouse cortical neuronal development.a Temporal expression patterns of Pogz mRNA in the brain (each n = 3). E embryonic day; wk week-old. b Regional expression pattern of Pogz mRNA in the E16.5 brain (each n = 3). VZ ventricular zone; SVZ subventricular zone. c Fluorescence in situ hybridization of coronal sections of the E16.5 brain for Pogz and immunostaining with antibodies against PAX6, SOX2, and TBR2. Scale bars, 100 µm. LV lateral ventricle; VZ ventricular zone; SVZ subventricular zone; IZ intermediate zone. d Impairment of neuronal migration by shRNA-mediated knockdown of Pogz in E18.5 mouse cortices electroporated at E14.5. CP cortical plate; IZ intermediate zone; SVZ subventricular zone. Scale bars, 50 µm. e Quantification of GFP+ cells in each layer (each n = 4). f GFP+ neurons were co-labeled for SATB2 (a layer II/III marker). g Quantification of GFP+ SATB2+ neurons (n = 4). h GFP+ neurons were co-labeled for CTIP2 (a layer V marker). i Quantification of GFP+ CTIP2+ neurons (n = 4). j GFP+ neurons were co-labeled for TBR1 (a layer VI marker). k Quantification of GFP+ TBR1+ neurons (n = 4). Note that Pogz knockdown had little effect on the proportion of SATB2+ GFP+, CTIP2+ GFP+ or TBR1+ GFP+ neurons and that the GFP+ cells with delayed migration were mostly SATB2+ neurons. h Same slice as in f. f, h, j Right panels, magnifications of the areas outlined with orange boxes. White scale bars, 50 µm; orange scale bars, 10 µm. a, b, g, i, k One-way ANOVA with Bonferroni–Dunn post hoc tests; a F10, 22 = 48.52; b F7, 16 = 73.23; g F2, 9 = 0.393; i F2, 9 = 1.079; k F2, 9 = 0.239. e Two-way repeated-measures ANOVA with Bonferroni–Dunn post hoc tests, F4, 27 = 39.28. ***P < 0.001. Data are presented as the mean ± s.e.m.