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. 2020 Oct 5;10:16490. doi: 10.1038/s41598-020-73752-3

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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 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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Expression of Spred2 mRNA in the lung of WT mice before and after BLM treatment. Lungs were harvested from untreated WT mice (naïve) or 1, 7 or 14 days after the intratracheal instillation of BLM. (a) The expression of Spred2 mRNA was examined on day 0 (naïve), 1 and 7 after BLM treatment by ISH. Left panels: H&E staining. Middle panels: ISH. Right panels: The area indicated by a rectangle in the middle panel is further magnified. Almost all cells expressed a significant level of Spred2 mRNA (blue-green dots) in naive mice as indicated by arrows. By contrast, Spred2 mRNA was expressed at a low level on day 1 and 7 as indicated by an arrow. The magnification of the original photos was 400×. The scale bars are 50 μm. (b) The expression of Spred2 mRNA on day 14 was examined by ISH. Top left panel: H&E staining. Bottom left panel: The area indicated by a rectangle in the top left panel is further magnified. Top right panel: ISH. Bottom right panel: The area indicated by a rectangle in the top right panel is further magnified. Spred2 mRNA accumulated in approximately 50% of bronchial epithelial cells, likely Clara cells by morphology, as indicated by arrows. The magnification of the original photos was 400×. The scale bars are 50 μm.