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. 2024 Feb 2;15:1275064. doi: 10.3389/fimmu.2024.1275064

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Copyright © 2024 Wu, Liu, Zhang, Ye, Gu, Jiang, Zhu, Ye, Li, Huang and Cao

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Validation of mRNA expression in IPF fibroblast model and human lung tissue. (A) Different mRNA levels of CALD1, CDH2 and POSTN in sequencing results. N1-N3: HLFs isolated from normal lung tissues adjacent to tumors; T1-T3: primary HLFs stimulated with recombinant human TGF-β1 (5 ng/ml) for 24 hours. (B) Comparison of CALD1, CDH2, POSTN mRNA expressions in HLFs between TGF-β1 group and normal control (NC) group. n=6 for each group. (C) mRNA expressions of CALD1, CDH2, POSTN in healthy control (HC) and IPF patients’ lung tissues measured by RT-qPCR. n=8 for each group. (D) Representative immunohistochemistry images and quantification (E) of CALD1, CDH2 and POSTN expression in lung fibrosis and normal lung tissues. n=8 for each group. Original magnification 200×, Scale bar 100 μm; Original magnification 400×, Scale bar 50μm. (F). RT-qPCR analysis of COL1A1 gene expression in lung fibroblasts treated with respective siRNA and TGF-β1. (G) Quantitative PCR analysis of ACTA2 gene expression in lung fibroblasts treated with respective siRNA and TGF-β1. n=3 for each group. (H) Representative western blot images and quantification of Collagen I and α-SMA expression in lung fibroblasts treated with respective siRNA and TGF-β1. *p< 0.05, **p < 0.001, ***p < 0.0001, and ****p < 0.00001, ns, not significant by Welch one-way ANOVA test, Error bars denote SD.