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. 2020 May 12;18:72. doi: 10.1186/s12964-020-00576-z

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — MLN4924-induced autophagy was a survival signal and ATF3 silencing enhanced drug toxicity via inducing apoptosis. a-b The proliferation inhibition by MLN4924 was significantly increased by simultaneously blocking autophagy with siBeclin1(a) or CQ(b). The combination of siBeclin1 or CQ with MLN4924 in EC1 and Kyse450 cells significantly increased proliferation inhibition by ATPLite assay. c-d Apoptosis induced by MLN4924 was significantly increased by simultaneously blocking autophagy with siBeclin1 or CQ. The combination of siBeclin1(c)or CQ (d) with MLN4924 in EC1 and Kyse450 cells significantly increased apoptosis by Annexin V /PI double staining. e Beclin1 knockdown increased cleaved PARP expression induced by MLN4924. EC1 and Kyse450 cells were transfected with control or Beclin1 siRNA for 48 h and then treated with 0.6 μmol/L MLN4924 for 48 h. Knockdown efficiency and cleaved PARP were assessed by IB analysis. f-h Blocking autophagy by siATF3 remarkably suppressed cell proliferation and induced cell apoptosis compared with MLN4924 alone. EC1 and Kyse450 cells were transfected with control or ATF3 siRNA for 48 h and then treated with 0.6 μmol/L MLN4924 for 48 h. Cell proliferation suppressed by MLN4924 was further significantly decreased by simultaneously silencing of ATF3 expression (f). Knockdown efficiency and cleaved PARP were assessed by IB analysis (g). Apoptosis detection by either annexin V and PI double staining (h)