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. 2020 Aug 26;10:14205. doi: 10.1038/s41598-020-69907-x

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© The Author(s) 2020

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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The coculture of HeLa/ADSC induces changes in the transcriptome of HeLa cells. (a) ADSCs were obtained from three donor patients. The table summarizes the main characteristics of the donor patients who underwent gastric bypass. (b) The purity of ADSCs in each patient was evaluated by flow cytometry analysis of cell surface markers including CD44, CD90, CD31 and CD45. The graph shows the percentage of positive and negative cells to each marker (n = 3 patients, 2 replicates, error bars = s.d.). (c) HeLa cells were cultured alone or in the presence of ADSCs by an indirect coculture system. The picture show that both cells lines are cultured in the same medium, but they are physically separated by a permeable membrane avoiding direct contact. (d) Bar chart shows the mRNAs altered in HeLa cells due to the presence of ADSCs. (e) The table shows the main molecular and cellular processes altered during coculture in HeLa. (f) The table shows the processes networks enriched during coculture in HeLa. (g) Schematic overview showing NF-kappa B as essential key hub driving gene expression, which was predicted to be activated in HeLa cells cocultured with ADSCs. (h) Top 20 differentially expressed genes in HeLa during coculture with ADSCs. (i–j). Quantification of gene expression by ddPCR showing the validation of mRNAs altered in HeLa by the presence of ADSCs obtained from patients (j) or ATCC (k). Graphs represent three biological replicates, and the error bars are s.d., *p < 0.05).