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. 2019 Mar 19;9:4868. doi: 10.1038/s41598-019-41057-9

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© The Author(s) 2019

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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TLP-mediated transcriptional shutdown after etoposide exposure is crucial for efficient apoptosis induction. (a,b) Control (ctrl) and TLP-knockdown (shTLP) HeLa cells were treated with 50 μM etoposide (Eto) for 24 h or left untreated (NT), and the total H2Bub level was monitored by Western blotting. Where indicated, α-amanitin was added to the final concentration of 2 µg/ml 12 h before analysis. (c) Control and TLP-knockdown HeLa cells were treated with 50 μM etoposide and indicated concentrations of α-amanitin. Cell viability was determined by SF assay and normalized to the cell number under nontreated conditions. The y-axis represents the average and S.D. of three independent experiments. (d) Control and TLP-knockdown HeLa cells were treated as in (b), and the conversion of Procaspase-3 (pro-Casp3) to Caspase-3 (Casp3) was monitored by Western blotting. (e) Etoposide sensitivity of B02-treated HeLa cells. Control and TLP-knockdown HeLa cells were treated with indicated concentrations of etoposide with or without 10 μM B02 for 36 h, and cell viability was determined by SF assay. The y-axis represents the average and S.D. of three independent experiments.