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. 2020 Jun 19;11:3111. doi: 10.1038/s41467-020-16947-6

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Schematic representation of conditional Nolz1 locus and Cre deleter lines used to ablate Nolz1 in the following regions: En1Cre for midbrain, FoxD1Cre for hypothalamic and FoxG1-IRES-Cre line for telencephalic selective recombination. b–e Immunostaining for NOLZ1 and GlycoDAT on coronal sections of E18.5 mouse striatum, hypothalamus and midbrain in the following mouse lines: Control (b–b′′), FoxG1-IRES-Cre;Nolz1 ^fl/fl (c–c′′), Foxd1Cre;Nolz1 ^fl/fl (d–d′′) and En1Cre;Nolz1 ^fl/fl (e–e′′) mutant embryos. Arrow in c′ points to DA axon bundles crossing the midline. f Representative outline of coronal sections from E18.5 mouse striatum (top), hypothalamus (middle) and midbrain (bottom) indicating regions represented in b–e. Cx (cortex), Str (striatum), MFB (medial forebrain bundle), SN (substantia nigra). g Graph showing GlycoDAT fibre density in the striatum of E18.5 Nolz1^−/−, FoxG1-IRES-Cre;Nolz1 ^fl/fl, FoxD1Cre;Nolz1 ^fl/fl and En1Cre;Nolz1 ^fl/fl mutant embryos relative to Wt embryos (n = 3 biologically independent samples). Wt values are normalized to 100%. Mean values ± standard deviation. Wt versus Nolz1^−/−: Two-sided, unpaired T-test ***p = 0.00098. Wt versus FoxG1-IRES-Cre;Nolz1 ^fl/fl: Two-sided, unpaired T-test **p = 0.00605. h Analysis of striatonigral and striatopallidal-specific markers in control and FoxG1-IRES-Cre;Nolz1 ^fl/fl mutant embryos by in situ hybridization. Coronal striatal sections of E15.5 embryos. i Downregulation of axon guidance markers in E15.5 in FoxG1-IRES-Cre;Nolz1 ^fl/fl mutant embryos as shown by in situ hybridization on coronal sections. j–k Reduction of DARPP32 expression and innervation by GlycoDAT labelled axons in the striatum of E18.5 FoxG1-IRES-Cre Nolz1 ^fl/fl mutant embryos visualized by immunohistochemistry on coronal sections. Data are representative of three independent experiments (b–e, h–k). Scale bar (b–k): 200 μm. Source data are provided as a source data file.