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. 2017 Nov 6;7:14579. doi: 10.1038/s41598-017-15074-5

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© The Author(s) 2017

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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Knockdown of NMI inhibits TNF-α plus CHX stimulated cell death. (A) HeLa-shCtrl and HeLa-shNMI cells were plated in 12-well plates. The following day, the cells were treated as indicated (TNF-α: 5 ng/ml, CHX: 5 μg/ml, IFNα:1000U/ml), and the cells were obtained and the ratio of cell death was determined with a Trypan blue staining assay. The results are representative of three independent experiments, and the error bars represent the SD.*p < 0.05. (B) H1299-shCtrl and H1299-shNMI cells were treated as in (A), and cell death was analysed by a Trypan blue staining assay. The results are representative of three independent experiments, and the error bars represent the SD.*p < 0.05, **p < 0.01. (C) FACS analysis. HeLa-shCtrl, HeLa-shNMI and HeLa-NMI cells were untreated or treated with IFNα for 12 h and then treated with TNF-α plus CHX for the indicated times (TNF-α: 5 ng/ml, CHX: 5 μg/ml). The cells were harvested, stained with propidium iodide (PI) and annexin V -FITC analyzed by flow cytometry. Data are represented as means ± s.e.m. P-values were calculated using Student’s t-test. *p < 0.05, **p < 0.01.