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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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Characterization of lungs from naïve WT and Spred2^−/− mice 1 day after BLM administration. Mice were injected intratracheally with 1.5 mg/kg BLM and lung tissues were harvested after 24 h. Tissue sections were subjected to H&E staining, TUNEL and IHC with anti-Ly6G Ab. To detect the proliferation of cells, BrdU was intraperitoneally injected 2 h before the harvest and the incorporation of BrdU was evaluated by IHC. Representative images are shown. The original magnification was 200 × for H&E staining and 400 × for TUNEL, Ly6G or BrdU staining. Arrows indicate positive cells. The number of TUNEL⁺ or BrdU⁺ cells in five bronchi were counted and presented as TUNEL⁺ or BrdU⁺ cells/100 cells. Ly6G⁺ cells in five high power field (400×) were counted and the results are presented as mean ± SEM. n = 5. Scale bars are 100 μm for H&E and Ly6G staining and 50 μm for TUNEL and BrdU staining.