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. 2022 Jul 18;13:4157. doi: 10.1038/s41467-022-31642-4

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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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Fig. 3 — a Venn diagram indicates upregulated genes that presented at least a twofold significant change in expression in MDCK-pTR GFP-Ras^V12 cells treated with CM of X-ray- (red) and oncogene-induced senescent IMR-90 cells (green) than in control cells. Out of 2518 genes detected in two samples, 247 were commonly upregulated in both. An enrichment test using GO analysis was performed with those commonly upregulated genes in MDCK-pTR GFP-Ras^V12 cells. The 15 most enriched terms within the biological process of GO are described in the bar graph. b The appearance of MDCK-pTR GFP-Ras^V12 cells treated with CM derived from oncogene- or X-ray-induced senescent IMR-90 cells for 24 h. Scale bar, 100 μm. c RT-qPCR analysis of EMT markers in treated MDCK-pTR GFP-Ras^V12 cells (H-Ras:^V12 CDH1, P < 0.001; ZO1, P = 0.0016; fibronectin, P < 0.001; ZEB1, P = 0.0018), (X-ray: CDH1, P < 0.001; ZO1, P < 0.001; fibronectin, P < 0.001; ZEB1, P = 0.044). d The appearance of nontreated MDCK-pTR GFP-Ras^V12 cells, MDCK-pTR GFP-Ras^V12 cells treated with CM derived from oncogene-induced senescent IMR-90 cells, X-ray-induced senescent IMR-90 cells, or cells treated with 20 ng/ml HGF for 24 h. Scale bar; 100 μm. e RT-qPCR analysis of EMT markers in MDCK-pTR GFP-Ras^V12 cells treated with 20 ng/mL HGF (CDH1, P < 0.001; ZO1, P < 0.001; fibronectin, P < 0.001; ZEB1, P < 0.001). CDH1 and ZO1 are shown as epithelial markers (c, e). Fibronectin and ZEB1 are shown as mesenchymal markers (c, e). Representative data from three independent experiments are shown. Error bars indicate the mean ± standard deviation of three independent measurements for all graphs. *P < 0.05, **P < 0.01, or ***P < 0.001 by the unpaired two-sided t test (c, e).