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. 2020 Aug 31;11:4363. doi: 10.1038/s41467-020-18175-4

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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. 3 — a–h Abhd4 mRNA is present exclusively in the ventricular zone along with the lateral (b, g) and third ventricles (c, h) at both E16.5 (a–d) and P1 (f–h) wild-type (+/+) mice. The specificity of the Abhd4 riboprobe is validated in Abhd4-knockout (−/−) animals (e). CP, cortical plate; IZ, intermediate zone; SVZ, subventricular zone; VZ, ventricular zone. High-power confocal imaging outlines the plasma membrane of ChR2-GFP-electroporated cells and delimits multi-color RNAscope analysis into single cells within the heterogeneous and densely packed cell layer of the ventricular zone. Abhd4 mRNA typically colocalizes with the radial glia progenitor cell marker Slc1a3 mRNA (encoding GLAST1 protein) (i), whereas other cells are often devoid of both markers (j). k Correlation analysis of Abhd4 mRNA levels with Glast1 mRNA levels in single cells (Spearman’s rank correlation, Abhd4/Glast1: R = 0.48, P < 0.0001; n = 92 cells from n = 4 mice). The scatter plot shows data from individual cells normalized to the median value of the respective mRNA levels. l, m Abhd4 mRNA distribution in attached daughter cells marked by PHH3-immunostaining. Arrows point to the mitotic cleavage furrow between the dividing cells. n Quantification of Abhd4 mRNA allocation within PHH3-positive daughter cells (Shapiro-Wilk normality test; W = 0.9792; P = 0.5439; n = 24 sections from n = 3 animals). o–q Representative images for Abhd4 in situ hybridization combined with TBR2-immunostaining. Abhd4 mRNA shows complementary distribution to TBR2 protein-containing intermediate progenitor cells. Scale bars: a: 100 μm, b–e, g–h, o–q: 50 μm, f: 500 μm, i, j, l, m: 2 μm. Source data are provided as a Source Data file.