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. 2019 Feb 5;9:1448. doi: 10.1038/s41598-018-37850-7

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© The Author(s) 2019

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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NeuroD2 is required for survival and terminal differentiation of MLIs. (A) Absence of PV⁺ MLI in the cerebellum of NeuroD1^−/− mutants. Fluorescent immunostaining of sagittal sections (lobule 5) at P15. Arrowheads point to PV⁺ MLIs in control mice. Scale bar, 50 µm. (B) Nuclear DAPI staining of sagittal sections (lobule 5) at P15. Scale bar, 50 µm. (C) Increased density of DAPI⁺ nuclei in the ML of NeuroD^−/− mutants. n = 4–5 per genotype. (D) Fluorescent immunostaining of sagittal sections (lobule 5) at P25. Arrowheads point to PV⁺ MLIs in control mice. Scale bar, 50 µm. (E) H&E staining and immunostaining for GAD67 on sagittal sections (P25, lobule 5). (F) Reduced density of GAD67⁺ interneurons in the ML of NeuroD2^−/− mutants. Quantification of H&E and GAD⁺ stainings in lobule 5, n = 3 per genotype.