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. 2021 Jan 27;12:612. doi: 10.1038/s41467-021-20902-4

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© The Author(s) 2021

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. 1 — a LWs (marked by dotted lines) with other forebrain tissues were subjected to X-gal staining. Scale bar, 500 μm. Cx cortex, CPu caudate putamen, ac anterior commissure. b The LWs were co-stained with ANKS1A, FOP, and CNTRL antibodies. The 3D SIM images were analyzed by the Imaris software. c 3DSIM micrographs of representative BBs. ANKS1A-VN was injected into the LV at E16.5 and subjected to in utero electroporation. The LWs at P4 were then co-stained for FOP and various BB markers with antibodies. Scale bar for b, c, 500 nm. d A median line and upper and lower quartile are presented in box and whisker plot of the axial distances of BB proteins from immature E1 cells. CEP164, n = 24; Centrin, n = 19; CNTRL, n = 14; ODF2, n = 15; ANKS1A-VN, n = 18. A green-shaded box marks a FOP-stained region brighter than 30% of the maximum intensity. e Table showing the radial distances of BB proteins from immature cells. The center of the FOP-positive ring image was used as a reference for the radial distances of BB proteins; n indicates the number of BBs used for statistical analysis. SD standard deviation. f Cartoon depiction of the BB proteins shown in d, e. g Whole-cell lysates (WCL) were prepared from six LWs at P4 and then the protein complexes were precipitated with the C-terminal specific antibody to detect the ANKS1A-associated proteins. h–j 3DSIM micrographs of BBs in E1 cells representing three different developmental stages. White arrowheads mark ANKS1A-VN present in FOP-stained SDAs. Scale bar for h–j, 500 nm. k Data shown in h–j were quantified with the total number of the double-positive FOP⁺CNTRL⁺ BBs set to 100%. Then the percentage of triple-positive ANKS1A⁺FOP⁺CNTRL⁺ BBs was calculated. Undifferentiated (UD), n = 67; immature (IM), n = 195; mature (M), n = 227 where n indicates the number of BBs analyzed.