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. 2018 Nov 9;9:4725. doi: 10.1038/s41467-018-07171-4

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© The Author(s) 2018

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. 7 — Ebf1 cKO affects iSPN migration and intermix with dSPN. a–j Distribution of FoxP2^high; Ctip2 + dSPN is altered in Ebf1 cKOs at E17.5, with lower density in the dorsal (Dor) and higher in the lateral (Lat) part of the striatum (Str); the two regions are defined in (j) (n = 3 for each condition). k In control Dor Str, 53 ± 3.5% of all Ctip2⁺ SPN are Foxp2⁺, 24 ± 6% (p = 0.0079) in Islet1^Cre;Ebf1^fl/⁻ and 21 ± 8%. (p = 0.03) in Dlx5/6::Cre;Ebf1^fl/⁻. In control Lat Str, Foxp2 + cells form 41 ± 11% of Ctip2 + SPN, raising to 54 ± 3% in Islet1^Cre;Ebf1^fl/⁻ (p = 0.01) and to 58 ± 11% in Dlx5/6::Cre;Ebf1^fl/⁻ (p = 0.03). l–t Distribution of Drd2-EGFP⁺;Ctip2⁺ iSPN is altered by Ebf1 cKO, with higher density in Dor Str and lower in Lat Str. u Quantification of iSPN distribution. In control Dor Str, 22 ± 12% of Ctip2⁺ SPN are GFP⁺. This percentage increases to 37 ± 11% in Islet1^Cre;Ebf1^fl/⁻;Drd2-EGFP (p = 0.007) and to 33 ± 7% in Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP (p = 0.03). Conversely, in the Lat Str, the percentage of of double-positive GFP⁺Ctip2⁺ iSPN equals 40 ± 6% in controls, 14 ± 6% in Islet1^Cre;Ebf1^fl/⁻;Drd2-EGFP (p = 0.008) and 13 ± 4% in Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP embryos (p = 0.008). v, w Analysis of iSPN migration at E15.5 in acute slices of Islet1^Cre;Ebf1^fl/⁻; Drd2-EGFP and Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP embryos. (v) Multidirectional displacement vectors from sample video tracking of control, Islet1^Cre;Ebf1^fl/⁻; Drd2-EGFP and Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP iSPN (colors illustrate directionality). w Trajectory analysis (Islet1^Cre;Ebf1^fl/⁻; Drd2-EGFP: 58 cells tracked, n = 3 independent experiments; Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP 29 cells tracked, n = 2 independent experiments) shows reduction of iSPN average speed after 180’ in Ebf1 cKOs (17,25 ± 1 μm/h for Islet1^Cre;Ebf1^fl/⁻; Drd2-EGFP, p = 0.02; 16,56 ± 1 μm/h for Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP p = 0.03) without significant alteration of maximum speed (117,8 ± 5 μm/h and 131 ± 6 μm/h, respectively; p = 0.47 for Islet1^Cre;Ebf1^fl/⁻; Drd2-EGFP and p = 0.55 for Dlx5/6::Cre;Ebf1^fl/⁻;Drd2-EGFP when compared to control iSPN). Results are presented as mean ± standard deviation (k, u) or s.e.m (w). Two-tailed non-parametric Mann–Whitney U test was used for statistical comparison. * indicates p-value < 0.05, ** p-value < 0.01. Scale bars equal 200 μm (a,d,g,l,o,r), 50 μm (b,c,e,f,h,I,m,n,p,q,s,t). Dor, Dorsal; Lat, lateral; Str, striatum