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. 2021 Jul 2;7(27):eabf1973. doi: 10.1126/sciadv.abf1973

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Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 4 — (A) Zeb2 represses Nrp1 expression. (B) ISH for Nrp1 in Zeb2^fl/fl and Zeb2^fl/fl Nex^Cre animals at E15.5. Scale bar, 50 μm. (C and D) Nrp1 and integrins mediate enhanced adhesion of Zeb2-deficient neurons. (C) Adherence of Zeb2^fl/fl primary cortical neurons transfected with scrambled short hairpin–mediated RNA (shScr), an Nrp1 shRNA (shNrp1) or ItgB1DN, and Cre as indicated to laminin-coated surfaces for 2 hours. Scale bar, 15 μm. (D) Lamellipodial spreading. N = 21 Zeb2^fl/fl + shScr, 26 Zeb2^fl/fl + Cre + shScr, 26 Zeb2^fl/fl Cre + shNrp1, and 22 Zeb2^fl/fl + Cre + ItgB1DN. One-way ANOVA (Kruskal-Wallis test) with Dunn’s multiple comparison test. (E to G) Zeb2 regulates laminar position through repression of Nrp1. (E) Zeb2^fl/fl animals were in utero electroporated at E14.5 with shScr, shNrp1, and Cre as indicated and analyzed at E18.5. (F) GFP⁺ neurons in the neocortex. Scale bar, 100 μm. (G) Laminar distribution of GFP⁺ neurons at E18.5. N = 11 Zeb2^fl/fl + shScr, 5 Zeb2^fl/fl + shScr and Cre, and 7 Zeb2^fl/fl + Cre + shNrp1 animals. Two-way ANOVA with Bonferroni post hoc test.