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. 2021 Jan 19;28(6):1941–1954. doi: 10.1038/s41418-020-00725-4

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 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 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. 4 — a KYSE150 cells were harvested 48 h after transfection and fractionated into nuclear (Nuc) and cytoplasmic (Cyto) components. The translocation of NF-κB were analyzed by measuring p65 subunit level in each component using WB. GAPDH and LaminB1 were employed as internal control, respectively. b Immunofluorescence microscopy for localization of p65 in KYSE150 cells 48 h after transfection. The nuclei were stained with 1 μg/ml DAPI (blue). Scale bar, 50 µm. c Transfected cells were treated with 10 μM IKK inhibitor BAY11-7082 for 12 h before nuclear/cytoplasmic fractionation. d KYSE150 cells with IKK knockdown were treated with BAY11-7082. Autophagic markers (LC3-II and p62) were examined to evaluate the selectively inhibition of BAY11-7082 on IKK. e Starvation induced apoptosis was analyzed by flow cytometry with Hochest33342/PI double staining. Apoptosis(%) = percentage of Hochest+ve/PI−ve cells. f Proliferation rates of transient transfected KYSE150 cells were revealed by crystal violet assay. Mock-transfected cells were used as control. *p < 0.05; **p < 0.01.