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. 2022 Oct 19;12:17440. doi: 10.1038/s41598-022-22156-6

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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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Loss of claudin-3 promoted lymphangiogenesis in primary tumor tissue. (a) Representative images of the lymphatic vessel marker LYVE-1 staining (green staining) in melanoma tissue of claudin-3^+/+ and claudin-3^−/− mice. Dashed lines indicate margin of tumor tissue. Left panel: 4X picture; scale bar, 1000 μm. Right panel: 20X picture; scale bar, 100 μm. (b) Statistical analysis of the lymphatic vessels under a high magnification (20X) fluorescence microscope. Values are shown as the mean ± SEM. (***p < 0.001) (n = 20 fields per group) (c) Representative images of CD31 staining (red staining) in melanoma tissue of claudin-3^+/+ and claudin-3^−/− mice. Left panel: 4X picture; scale bar, 1000 μm. Right panel: 20X picture; scale bar, 100 μm. (d) Statistical analysis of the blood vessels under a high magnification (20X) fluorescence microscope. Values are shown as the mean ± SEM. (ns: no significance) (n = 20 fields per group) (e) Co-staining of tumor tissue from the claudin-3^+/+ and claudin-3^−/− mice with anti-LYVE-1 antibody (green staining) and anti-claudin-3 antibody (red staining). Scale bar, 10 μm. (f) Quantification of claudin-3 fluorescence intensity on LYVE-1 + lymphatics in e. (***p < 0.001) (n = 24 vessels per group).