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. 2019 Sep 24;10(10):709. doi: 10.1038/s41419-019-1872-y

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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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Fig. 6 — a CP-dn-ATF5 reduces expression of over-expressed FLAG-survivin, but to levels far in excess of endogenous survivin. T98G cultures were infected with lentivirus expressing either pLVX-EF1α-IRES-mCherry (pLV in Figure) or pLVX-EF1α-FLAG-survivin-IRES-mCherry (pLV-survivin in Figure) as indicated and 24 h later treated with or without 100 µM CP-dn-ATF5 for 3 d and assessed for exogenous and endogenous survivin by western immunoblotting with anti-survivin. Long exposure shows levels of endogenous survivin. b Survivin over-expression does not rescue the effects of CP-dn-ATF5 on number or appearance of T98G cells. Cultures were infected with lentiviruses described in panel a as indicated and 24 h later were treated with 100 µM CP-dn-ATF5 for 3 days. Panels show immunofluoresence for mcherry (red) or phase contrast images. Scale bars = 50 µm. c Survivin over-expression does not rescue T98G cells from apoptotic death promoted by CP-dn-ATF5. Cultures were infected with lentivirus described in panel a and 24 h later were treated with 100 µM CP-dn-ATF5 for 3 days. Cultures were harvested and analyzed for proportion of apoptotic cells by flow cytometry. Left panel shows the representative cytometry data and right panel shows quantitative results from three independent experiments, each in triplicate. d Survivin over-expression does not rescue cell number in T98G cultures treated with CP-dn-ATF5. Cultures were infected with above described lentiviruses and 24 h later were treated with or without 100 µM CP-dn-ATF5 as indicated for 3 days. Cultures were harvested and analyzed for total cell numbers. Data are from three independent experiments, each in triplicate. e Survivin over-expression does not rescue promotion of apoptosis by FLAG-GFP-dn-ATF5 in multiple cancer cell lines. Indicated cell lines were co-transfected as shown with a ratio of pLVX:pLE constructs of 3:1 and transfected (GFP+) cells were assessed for proportion with apoptotic nuclei 3 days later. Data are from three independent experiments