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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. 6 — a Confocal microscopic analysis of LWs from littermates at 22 months of age (top panels). A representative cell (marked by yellow arrowheads) in each panel is enlarged to reveal motile cilia (green), BBs (red) (middle panels), and F-actin (gray) at the apical and subapical levels (bottom panels). The cell border and actin network around a BB patch are outlined with white and red dotted lines, respectively. b An SEM analysis of LWs from littermates at 22 months of age. Scale bar for a, b, 10 μm. c–e Data from a were quantified. The mice used for this quantification ranged from 18 to 22 months of age. For c: ANKS1A^+/+ set, n = 110 cells from 4 mice; ANKS1A^−/−, n = 113 cells from 4 mice. For d, e: ANKS1A^+/+ group, n = 87 cells from 4 mice; ANKS1A^−/−, n = 85 cells from 4 mice. f 3DSIM micrographs of LWs depict triple staining with FOP, ODF2, and CNTRL. White boxes indicate the regions of ×3 magnification that reveal SDA staining with each antibody (second and third panels). Scale bar, 10 μm. g Representative SDA images (top) together with their cartoon depiction (bottom). Scale bar, 200 nm. h The central angles were calculated. ANKS1A^+/+ set, n = 164 BBs from 3 mice; ANKS1A^−/−, n = 160 BBs from 3 mice. i Data in f were quantified. Data represent mean ± SD. ANKS1A^+/+ group, n = 50 cells; ANKS1A^−/−, n = 56 cells. j, k A ribbon of consecutive sections (50 nm) was placed on a one-hole grid and subsequently analyzed by TEM. The resulting micrographs were aligned to measure the overall BF size. Data represent mean ± SD. ANKS1A^+/+ set, n = 50 BBs; ANKS1A^−/−, n = 54 BBs. Scale bar, 100 nm.