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. 2020 Apr 10;10:6224. doi: 10.1038/s41598-020-63085-6

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© The Author(s) 2020

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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Particulate matter from heavy diesel engine disturbs mucus secretion from the mucociliary epithelium in B. orientalis and X. laevis embryos. (A) Increasing doses of PM were administered to B. orientalis embryos and secreted mucus levels were analyzed by ELLA using WGA-HRP. 50 µg/mL or 100 µg/mL of PM significantly disturbed mucus secretion from the embryonic epidermis. Statistical analysis was performed using one-way ANOVA. From left to right, n = 21, 16, 20. (B) The WGA positive mucus vesicles were visualized by fluorescent microscopy. PM exposure significantly accumulated mucus vesicles in the cytoplasm. Scale bar = 10 μm. (C) Quantification of WGA positive mucus vesicles in (B). Statistical analysis was performed using one-way ANOVA. From left to right, n = 4, 4, 4. (D) Particulate matter-induced hyposecretion of mucus was recovered by α-tocopherol in Xenopus embryos. Statistical analysis was performed using one-way ANOVA. From left to right, n = 24, 22, 22. (E) The WGA positive mucus vesicles were visualized by fluorescent microscopy. The exposure to the particulate matter significantly accumulated the mucus vesicles in the cytosol. Scale bar = 20 μm. (F,G) Quantification of WGA positive mucus vesicles in (E). For (F), statistical analysis was performed using one-way ANOVA. From left to right, n = 4, 4, 4. For (G), statistical analysis was performed using one-way ANOVA. From left to right, n = 30, 30, 30. Asterisks represent ***(p < 0.001), **(0.001 < p < 0.01), *(0.01 < p < 0.05), ns (0.05 < p). PM, particulate matter; ELLA, enzyme-linked lectin assay.