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. 2023 Feb 6;14:643. doi: 10.1038/s41467-023-36314-5

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© The Author(s) 2023

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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Fig. 1 — a Migration of human primary lung fibroblasts (pHLF) post-TGF-β1 induction for 24 h, with significantly more myofibroblast cells appearing than in mock- and inhibitor(s)-treated wells. Data were analyzed using a one-way ANOVA followed by a Dunnett’s post hoc test unless otherwise specified: TGF-β1 vs TGF-β1/TH5487 (P = 0.0001); TGF-β1 vs TGF-β1/Dex (P = 0.6954); TGF-β1 vs TGF-β1/Nin (P = 0.0001); TGF-β1 vs TGF-β1/Veh (P < 0.0001). ns: not significant. Source data are provided as Source data file. Data are representative of 4 independent experiments containing 3 biological replicates (scale bar = 180 μm). Data are presented as the mean ± standard error of the mean (a, c, e, f). b Immunostaining of pHLF cells (green) following 24 h of TGF-β1 treatment, with TH5487 treatment displaying visually reduced levels of collagen (COL1A1), fibronectin (FN), vimentin (VIM), α-smooth muscle actin (α-SMA). Scale bar = 50 μm. Results shown from 3 independent experiments. c Migration of pHLF post-TGF-β1 induction for 24 h, and siRNA transfection targeting OGG1 or scrambled sequence (control). Data were analyzed using a one-way ANOVA followed by a Dunnett’s post hoc test: si Control/TGF-β1 vs si OGG1/TGF-β1 (P = 0.0008); si Control/TGF-β1 vs si OGG1/vehicle and si Control/TGF-β1 vs si Control/vehicle (P < 0.0001). Data are representative of 4 independent experiments containing 3 biological replicates (scale bar = 180 μm). Data are presented as the mean ± standard error of the mean. d Stress fiber formation was seen in response to TGF-β1 in Ogg1^+/+, but not Ogg1^−/− MF cells (left panel, F-actin). Expression and colocalization of α-SMA (red) with F-actin (green) are shown (right panels, scale bar = 90 μm). Results shown from 3 independent experiments. e The effects of TH5487 on transcription of α-Sma, Fn1, Vim, and Col1A1 in TGF-β1-stimulated MF cells as determined by qRT-PCR. TH5487 (10 μM), significantly decreased mRNA levels of all genes with compiled data representative of 3 independent experiments. Data were analyzed by one-way ANOVA followed by a Dunnett’s post hoc test. f TGF-β1-induced expression of αSma, Col1a1, Fn1, and Vim was decreased in Ogg1^−/− but not in Ogg1^+/+ MF cells. Data representative of 3 independent experiments. g Immunoblot analysis of TGF-β1-stimulated pHLF cells showing decreased levels of α-SMA, COL1A1, FN1, and VIM levels in OGG1-depleted cells by siRNA. TGF-β1, 2 ng/mL; TH5487, 10 μM; Nintedanib (Nin), 10 μM; Dexamethasone (Dex), 10 μM. Results shown from 3 independent experiments.