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. 2021 Jan 21;7:626140. doi: 10.3389/fmed.2020.626140

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Copyright © 2021 Xu, Li, Wang, Huang, Zhang, Zhang, Gu, Yu and Jia.

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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Cilomilast reduced the TGF-β1-induced profibrotic response in NRK-49F cells. NRK-49F cells stimulated with TGF-β1 were treated with cilomilast. (A) Cell viability was analyzed by CCK8 assay after treatment with celastrol for 24 h at increasing concentrations from 5 μM to 40 μM. (B) qRT-PCR analyses of Fn, Col-I, Col-III, and α-SMA mRNA expression (n = 3). (C) Western blot analyses of FN and Col-I protein expression in NRK-49F cells. (D) Densitometry of the western blot results in (B). (E) Representative images of immunofluorescence staining for FN in NRK-49F cells. (F) qRT-PCR analyses of Il-1β, Il-6, and Mcp-1 mRNA expression (n = 3). The data are presented as the mean ± SEM. Statistically significant differences were determined by one-way ANOVA and two-way ANOVA. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, ^####P < 0.0001, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.