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. 2018 Apr 18;28(7):2577–2593. doi: 10.1093/cercor/bhy082

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© The Author(s) 2018. Published by Oxford University Press.

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

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Figure 7. — Unaltered number and density of intermediate progenitors or in their expression of Cxcl12/Sdf1 in E13.5 Tie2Cre;Vegfa^fl/fl mutant forebrains. (A) Images of the dorsal cortex of E13.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant forebrains immunostained for Tbr2⁺ intermediate progenitor cells (IPCs) and with the vasculature labeled with isolectin-B4. Spots superimposed on Tbr2⁺ nuclei show automated detection of Tbr2⁺ cells using Imaris software spot module function, with spots in bottom panels showing the distribution of Tbr2⁺ cells and surface rendering of the vasculature used in quantifications shown in bar charts. (B) Graphs show the total number (left) and density of Tbr2⁺ cells within the neurogenic basal VZ/SVZ and normalized for differences in cortical thickness in E13.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant forebrains (second left). Graphs on right shows the surface area of IB4⁺ blood vessels in the cortex of E13.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant measured from surface renderings generated with Imaris software and the mean intra-cellular distance of Tbr2+ cells. (C) In situ hybridization analysis of Cxcl12/Sdf1 transcripts in the E13.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant forebrains. (D) Immunolabelling of Tbr2⁺ IPs (top panels), Ctip2⁺ and Satb2⁺ pyramidal neurons (bottom panels) in the dorsal cortex of E17.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant forebrains with (E) bar charts showing the mean total numbers and intercellular distances of Ctip2⁺ and Satb2⁺ cells. (F) Expression of Cxcl12/Sdf1 transcripts in the E17.5 Vegfa^fl/fl control and Tie2Cre;Vegfa^fl/fl mutant forebrains (t-test, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) (Bar graphs show mean values ±SEM).