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. 2021 Nov 4;11:21642. doi: 10.1038/s41598-021-01004-z

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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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Claudin-9 is a component of FS cells. (a) RT-PCR of the pituitary gland of adult male and female mice. Since the forward and reverse primers were designed in the same exon, the genome DNA was used as a positive control (PC) and water was used as a negative control (NC). The reaction mix without reverse transcriptase (RT) also served as a negative control. (b) Real-time RT-PCR of the adult mouse pituitary gland. The expression levels were normalized using known concentration of DNA fragments as standards. The expression level of cldn9 was set to 1. Error bars indicate s.d. (c) Immunoprecipitation of claudin-9 from adult mouse organs using rat anti-claudin-9 (clone 1B1) and immunoblotting using goat anti-claudin-9 pAb. Human CLDN9-expressing HEK293T cell lysate served as a positive control (PC). Note that the claudin-9 band was detected in the pituitary gland lysate as well as in the inner ear and olfactory mucosa. (d), (e) Immunostaining of the adult mouse anterior pituitary gland using anti-claudin-6 pAb, anti-claudin-7 pAb, anti-claudin-8 pAb, anti-claudin-9 mAb (clone 1B1), or anti-claudin-12 pAb together with anti-occludin mAb or pAb and DAPI.