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. 2021 May 25;11:10931. doi: 10.1038/s41598-021-90263-x

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© The Author(s) 2021

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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Anti-fibrosis effects of hit compounds in phenomic-based 2D assay of endothelial cells (HUVECs). (A) Intensity of α-SMA and F-actin in HUVEC cells in 2D culture system treating with TGF-β1. Cells were treated with 2.5, 5, 10, or 20 ng/ml TGF-β1 (B) Calculated intensity of α-SMA and F-actin when HUVEC cells were treated with TGF-β1 concentration. (C) α-SMA expression (left panel) and the quantitative of western blot image was analyzed (right panel) depending on treating with TGF-β1 concentration in HUVEC cells. (D) Representative images of α-SMA treated with 10 µM of pirfenidone, 0.5 µM or 1 µM of retinoic acid and forskolin with 20 ng/ml TGF-β1 for 48 h in HUVEC cells. (E,F) Expression of α-SMA when HUVEC cells were treated with 0.5 or 1 µM (E) Retinoic acid or (F) Forskolin. All images were obtained using the Operetta CLS system. Data are expressed as means ± SD (n = 3). ^*P < 0.05 and ^**P < 0.01 compared to the control group, ^#P < 0.05 and ^##P < 0.01 compared to the TGF-β1 treatment group.