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. 2020 Feb 4;10:1804. doi: 10.1038/s41598-020-58642-y

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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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TNF-α increases BCSCs and TAZ expression in MCF-7 breast cancer cells. (A) TNF-α increases BCSCs in MCF7 cells, as measured by mammosphere culture. MCF7 cells were treated with 10 ng/ml TNF-α or 0.1% BSA for 48 h, and cells were seeded at a density of 1,000 cells per well and cultured for 14 d. The number of primary mammospheres with diameter ≥60 μm was then quantified. (B) The number of primary mammospheres per 1,000 cells initially seeded was quantified (mean ± SEM; n = 3). **P < 0.01, t-test. (C) TNF-α increases BCSCs in MCF7 cells as measured by ALDH assays. MCF7 cells were treated with 10 ng/ml TNF-α or 0.1% BSA for 48 h. Cells were collected for aldehyde dehydrogenase assays by FACS. (D) The percentage of ALDH + cells was quantified (mean ± SEM; n = 3). *P < 0.05, t-test. (E) TNF-α increases BCSCs in MCF7 as measured by CD marker staining. MCF7 cells were treated with 10 ng/ml TNF-α or 0.1% BSA for 48 h. The cells were collected for CD marker staining by FACS. The percentage of CD44 + cells was quantified (mean ± SEM; n = 3). **P < 0.01, t-test. (F) TNF-α induces TAZ protein expression in MCF7 cells. MCF7 cells were treated with 10 ng/ml TNF-α or 0.1% BSA for 24 h and 48 h, respectively. TAZ protein levels were measured by Western blotting. (G) TNF-α induces TAZ and CYR61 mRNA expression in MCF7 cells. TAZ and CYR61 mRNA levels were analyzed by RT-qPCR. Data are normalized to untreated samples (mean ± SEM; n = 3). **P < 0.01, t-test. NS indicates not significant.