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. 2020 Mar 30;13(3):dmm042515. doi: 10.1242/dmm.042515

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© 2020. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 1. — Arx^(GCG)10+7 expansion mutants exhibit age-dependent increase in non-cell-autonomous apoptosis. (A-E) Representative images of a parasagittal cortical section of P7 Arx^(GCG)10+7 showing CC3-immunoreactive (CC3⁺) (green) and Arx-immunoreactive (Arx⁺) (red) cells in the neocortex. (B-E) Enlarged images of the white box in A, showing DAPI (B), Arx (C), CC3 (D) and merged (E) immunostaining. (E) Open arrowhead shows a CC3⁺ cell and filled arrowhead shows a CC3⁺ cell with Arx⁺ nucleus. (F,G) Histograms comparing mean CC3⁺ cells at P1, P4, P7, P9 and P14 from cortices of WT and Arx^(GCG)10+7 littermates (N=8-15 WT, N=8-16 mutant) in neocortex (F) and hippocampus (G). (F) In neocortex, two-way ANOVA yields an effect of genotype [F(1,119)=5.338, P=0.0025]. (G) In hippocampus, two-way ANOVA yields no effect of genotype [F(1,72)=0.9080, P=0.2035]. (H) Mean fluorescein isothiocyanate (FITC)⁺ (TUNEL⁺) cells in neocortex from TUNEL assay at P7 (P=0.0051, unpaired Student's t-test, N=8 WT, 8 Arx). (I) Histograms comparing mean CC3⁺ cells that co-express Arx (Arx⁺ CC3⁺). Two-way ANOVA yields no effect of genotype [F(1,88)=0.2374, P=0.6273] (N=8-12 WT, 8-12 mutant). (J) Histograms comparing mean co-labeled (CC3⁺ Arx⁺) cells normalized to Arx⁺ density. Two-way ANOVA yields no effect of genotype [F(1,88)=0.01996, P=0.8880] (N=8-12 WT, 8-12 mutant). (K) Mean Arx⁺ density (Arx⁺ cells/mm²) at P7 in the retrosplenial agranular region (RSA) (P=0.0225) and motor cortex (P=0.0017) (unpaired Student's t-test) of Arx^(GCG)10+7 mutants compared to WT. F-G and I-J utilized two-way ANOVA with Bonferroni's multiple comparisons test. Scale bars: 500 µm (A) and 100 µm (E), with images in B-E scaled similarly. Means±s.e.m. are displayed, with exact N values displayed on graphs.*P<0.05, **P<0.01. ns, not significant.