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. 2015 Dec 15;29(24):2617–2632. doi: 10.1101/gad.268086.115

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© 2015 Hernandez-Enriquez et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 6. — Precrossing axon guidance defects from floor plate-derived Nrp2-deficient animals are rescued by inhibition of PlxnA1 signaling in vivo. (A–C) Representative confocal images of E11.5 spinal cord sections taken from littermates treated with TM (+TM) with the following genotypes: Nrp2^f/f;FoxA2^+/+;PlxnA1^+/− (A), Nrp2^f/f;FoxA2^+/CreERT2;PlxnA1^+/+ (B), and Nrp2^f/f;FoxA2^{CreERT2/CreERT2};PlxnA1^−/− (C). All transverse sections were processed for immunocytochemistry for Nrp2 (blue), Robo3.1 (red), and GFP (green). White arrowheads illustrate Nrp2-positive dorsal commissural axons projecting toward the floor plate (FP) and ventral commissure (VC) in Nrp2^f/f;FoxA2^+/+;PlxnA1^+/− (control) and Nrp2^f/f; FoxA2^{CreERT2/CreERT2};PlxnA1^−/− (rescued) embryos but missing in Nrp2^f/f; FoxA2^+/CreERT2;PlxnA1^+/+ animals. Bar, A–C, 125 µm. (D) Quantifications of Robo3.1-normalized fluorescence in Nrp2^+/f;FoxA2^+/+;PlxnA1^+/− +TM, Nrp2^f/f;FoxA2^+/CreERT2;PlxnA1^+/+ +TM, and Nrp2^f/f;FoxA2^{CreERT2/CreERT2};PlxnA1^−/− +TM littermate embryos. Data are means ± SEM from five to eight sections per embryo, where n = 3 embryos per genotype combination analyzed. ANOVA followed by post-hoc Tukey test, (*) P < 0.05 compared with Nrp2^+/f;FoxA2^+/+;PlxnA1^+/− +TM; (#) P < 0.05 compared with Nrp2^f/f;FoxA2^+/CreERT2;PlxnA1^+/+ +TM; no significant difference for comparison between Nrp2^+/f;FoxA2^+/+;PlxnA1^+/− +TM and Nrp2^f/f;FoxA2^{CreERT2/CreERT2};PlxnA1^−/− +TM. (E) Quantifications of normalized ventral commissure thickness in Robo3.1-positive axons in Nrp2^+/f;FoxA2^+/+;PlxnA1^+/− +TM, Nrp2^f/f;FoxA2^+/CreERT2;PlxnA1^+/+ +TM, and Nrp2^f/f;FoxA2^{CreERT2/CreERT2};PlxnA1^−/− +TM littermates. Data are means ± SEM from five to eight sections per embryo, where n = 3 embryos per genotype combination analyzed. ANOVA followed by post-hoc Tukey test, (*) P < 0.05 compared with Nrp2^+/f;FoxA2^+/+;PlxnA1^+/− +TM. (F) Schematic diagram of in vivo mouse genetics and the corresponding phenotypes observed.