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. 2022 Sep 29;12:16305. doi: 10.1038/s41598-022-20177-9

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© The Author(s) 2022

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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Identification of ERβ in human conjunctival goblet cells cultured from males and females. RNA was isolated from cultured goblet cells and RT-PCR performed using primers for human Erβ. A single band was detected at around 100 bp in 2 out of 4 samples from males (M) and 2 out of 3 samples from females (F) (A). Protein samples were collected from cell pellets and Western Blot analysis was performed using antibody against ERβ. Two bands at 100 and 75 kDa respectively were detected in female samples (B). Trace immunoreactivity of the same size as in females was present in male samples (B). Immunofluorescence microscopy was performed on cultured goblet cells using antibodies to ERβ (C and D). (C) indicate immunofluorescence to ERβ (red) in male cells; (D) indicate immunofluorescence to ERα (red) in female cells; an overlay of anti-ERβ, HPA-1 (green, indicates the secretory granules of goblet cells), and DAPI (blue, indicates cell nuclei) is shown next to the single channel image of ERβ. Magnification was × 200 in the upper panels; × 1000 in the lower panels. Original blots/gels are presented in Supplementary Figs. 5 and 6.