Fig. 2: Pogz-deficient mice show growth delay, changes in brain size, and altered neurogenesis. | Nature Communications

Fig. 2: Pogz-deficient mice show growth delay, changes in brain size, and altered neurogenesis.

From: Pogz deficiency leads to transcription dysregulation and impaired cerebellar activity underlying autism-like behavior in mice

Fig. 2

a Comparison of the body size of Pogz cKO−/− and control mice littermates (P11). b Body weight in four time points (P13, P21, P40, P60). Repeated measures ANOVA; n = 16 control (blue), 7 cKO+/− (gray), 7 cKO−/− (red), P = 1.1 × 10−15. c Brain weight at P11. n = 5 control, 5 cKO+/−, and 3 cKO−/−. d Coronal sections of embryonic cerebral cortex (E15.5) stained with anti-Pax6 and Tbr2 antibodies. Scale bar = 50 μm. VZ ventricular zone, SVZ subventricular zone. e, f Quantification of the thickness of the Tbr2+ and Pax6+ layers. Values are proportion relative to control mice. n = 5 control, 9 cKO+/−, 9 cKO−/− in four independent experiments. g, h Coronal sections of cerebral cortices (E15.5) stained with DAPI and antibody against phosphorylated histone 3 (pHH3). Scale bar = 100 μm. h Quantification of the number of the pHH3+ cells. Values are proportion relative to control mice. n = 5 control, 9 cKO+/−, 11 cKO−/− in two independent experiments. i, j Effects of Pogz-silencing on cell-cycle exit. i Differentiated neurons are EdU/GFP-double-positive while EdU/Ki67/GFP-triple-positive cells maintain progenitor potency. Arrowheads indicate triple-positive cells. Scale bar = 10 µm. At the bottom right is the scheme of the cell-cycle exit assay. j Quantification of EdU/Ki67/GFP-triple-positive cells among GFP/EdU double-positive ones (Cell exit index). Two sections were used per brain (more than 1600 cells were analyzed in each condition). n = 10 control, 9 sh-Pogz#1 (sh#1), 10 sh-Pogz#2 (sh#2) from two in utero independent experiments. k−m Reduced adult neurogenesis associated with Pogz deficiency. Quantification of either k DCX-positive cells, l Ki67-positive cells, m BrdU-positive cells in the dentate gyrus, 21 days post injection. Numbers are proportion relative to control mice. n = 6 control, 6 cKO+/−, 3 cKO−/−. The P values at the top of the plots are for the association between the quantitative measurements and the number of intact Pogz alleles calculated with a linear regression model. Pairwise comparisons between genotypes was calculated by two-tailed t test and the significance is represented by: *P < 0.05; **P < 0.01; ***P < 0.001; n.s. not significant. Quantitative data are mean ± SEM.

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