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. 2021 Dec 1;11:23256. doi: 10.1038/s41598-021-02420-x

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Blocking peptide WPSGGQMT abolishes 596 immunostaining at the bases of cilia. (a) Well differentiated HBEs were co-immunostained using 596 and centrin-2 antibodies without (top row) or with (bottom row) a blocking peptide corresponding to the 596 epitope. White arrows indicate ciliated cells. Zoomed-in images are shown at extreme right. (b) Rootletin isoforms containing a sequence homologous to the 596 epitope in CFTR. (c) Cartoon showing location of the pseudo-epitope in rootletin isoforms. (d) Immunoblot of well differentiated HBE cell lysates from a non-CF donor and six CF patients probed using rootletin antibody. Green and blue labels indicate donor is homozygous for F508del and a Class I mutation, respectively. Short (1) and long (2) exposures are shown. After short exposure note that rootletin was only detected in 596+ cell lysates. (e) Summary of rootletinX1 protein expression. Each point is the lysate from a different culture prepared from five non-CF donors, five 596+ donors and three 596- donors. Note that rootletinX1 band intensity is significantly lower in cells that are not immunostained using CFTR antibody 596 (mean ± SE, n = 16–17, **p = 0.0018, one-way ANOVA).