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. 2022 Dec 20;55(1):100–111. doi: 10.1038/s41588-022-01252-3

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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 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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Extended Data Fig. 9 — a&b, Immunofluorescence staining for α-SMA and 5hmC on paraffin sections of mouse lungs after intraperitoneal injections (IP) and intranasal (IN) challenges of PBS or Aspergillus fumigatus (ASP) (a). Scale bar: 50 μm. DNA was stained by DAPI. Arrows indicate 5hmC-positive SMCs. Inserts depict enlarged images of 5hmC-stained SMCs from the respective panels. MFIs of 5hmC signals in lung SMCs were quantified by Image J and are shown in b (PBS: n = 4, ASP IP: n = 5; two-tailed unpaired t-test: *P = 0.0126). c&d Immunofluorescence staining for α-SMA and 5hmC on paraffin sections of mouse lung after subcutaneous injections (SC) and IN challenges with ASP or PBS (c). Scale bar: 50 μm. DNA was stained by DAPI. Arrows indicate 5hmC-positive SMCs. Inserts depict enlarged images of 5hmC-stained SMCs from the respective panels. MFIs of 5hmC signals in lung SMCs were quantified by Image J and are shown in d (n = 5 independent animals; two-tailed unpaired t-test: **P = 0.0035). e&f, Immunofluorescence staining for α-SMA and 5hmC on lung paraffin sections from human donor (n = 4), COPD (n = 5) or cystic fibrosis (CF) patients (n = 5). Scale bar: 50 μm. DNA was stained by DAPI. Arrows indicate 5hmC-positive SMCs. Inserts depict enlarged images of 5hmC-stained SMCs from the respective panels. MFIs of 5hmC signals were quantified by Image J and are shown on the right (one-way ANOVA with Tukey’s post hoc test). g&h, Immunofluorescence staining for α-SMA and 5hmC on lung paraffin sections from mice exposed to normoxia and chronic hypoxia (10%) for 4 weeks (n = 5 independent animals). Scale bar: 50 μm. DNA was stained by DAPI. Arrows indicate 5hmC-positive SMCs. Inserts depict enlarged images of 5hmC-stained SMCs from the respective panels. MFIs of 5hmC signals were quantified by Image J and are shown on the right (n = 5; two-tailed unpaired t-test). The enlarged images in (a, c, e, g) were scaled to 150×150 pixels. Data in (b, d, f, h) are presented as mean values.

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