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. 2017 Apr 17;12(4):e0175888. doi: 10.1371/journal.pone.0175888

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© 2017 Teoh et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 5 — (A) The layering order of UL neurons (Cux1⁺) and DL neurons (Tbr1⁺) was maintained in Arfgef1^geo/geo but the IZ thickness reduced at E17.5. Scale bars, 200 μm. n = 3 per group. (B) At E17.5, the numbers of Tbr1⁺, Cux1⁺ and DAPI⁺ cells within a 200 μm cortical width starting from the edge of the lateral ventricle were counted. Scale bars, 200 μm. n = 4 per group. (C) The thickness of the CP (***p = 0.0003) and IZ (***p = 1.25E-06) in Arfgef1^geo/geo brains were significantly reduced. n = 5 per group. (D) In Arfgef1^geo/geo neocortex, the total Tbr1⁺ (*p = 0.02), Cux1⁺ (*p = 0.04) and DAPI⁺ (***p = 0.0007) cells per 200 μm cortical width significantly reduced compared to controls. Tbr1⁺, n = 4 per group; Cux1⁺, n = 4 per group; DAPI⁺, n = 8 per group. (E) In Arfgef1^geo/geo neocortex, DAPI⁺ cells accumulated in Bin 3 (*p = 0.02), Bin 4 (*p = 0.04), Bin 5 (*p = 0.03), Bin 6 (*p = 0.03) at the expense of DAPI⁺ cells in Bin 7 (***p<0.0001), and Bin 8 (*p = 0.01). n = 8 per group. (F) In the Arfgef1^geo/geo neocortex, Tbr1⁺ cells accumulated in Bin 6 (*p = 0.01) and at the expense of Tbr1⁺ cells in Bin 7 (**p = 0.004) and Bin 8 (*p = 0.03). n = 4 per group. (G) The Cux1⁺ cells accumulated in Bin 7 (*p = 0.01) and Bin 8 (*p = 0.03) at the expense of Cux1⁺ cells in Bin 9 (*p = 0.02). n = 4 per group. (H) Pregnant dams at E12.5 were injected with BrdU and the migration of cells in neocortex was quantified at E15.5. The percentage of BrdU⁺ cells in the VZ/SVZ (**p = 0.007) and IZ (*p = 0.04) significantly increased in Arfgef1^geo/geo compared to control brains. However, the percentage of BrdU⁺ cells in CP (p = 0.31) did not decrease. Arfgef1^geo/geo, n = 2; controls, n = 4. For all bar graphs, Student’s T-test was used for comparisons between groups. Data are shown as the mean ± SD. Bo, IZ-CP boundary; CP, cortical plate; IZ, intermediate zone; L1-L6, layers 1–6, MZ, marginal zone; VZ/SVZ, ventricular zone and subventricular zone.