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. 2018 May 25;8:8102. doi: 10.1038/s41598-018-26496-0

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© The Author(s) 2018

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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Bim mediated the efficacy of I-BET762 and gemcitabine (GEM) action in PDAC cells. (A) Panc-1 cells were treated with I-BET762 at indicated concentration, indicated protein level were analyzed by western blotting. (B) Panc-1 cells transfected with si control or si PUMA were treated with the combination of 1 μM I-BET762 and 5 μM GEM. Apoptosis was detected by flow cytometry. (C) Indicated cell lines were treated with 1 μM I-BET762 for 24 h. Bim mRNA level was analyzed by real-time PCR. (D) Indicated cell lines were treated with 1 μM I-BET762 or 5 μM JQ1 for 24 h, and Bim protein level was analyzed by western blotting. (E) Bim protein level in Bim-KO cells was analyzed by western blotting. (F) The distance migrated by WT and Bim-KO Panc-1 cells after treatment was quantified. The migrated distance was quantified by measuring the difference at time 0 and 24 h and was normalized to control. (G) I-BET762 at 1 μM inhibits the invasion of WT and Bim-KO Panc-1 cells. The invaded PDAC cells were quantified by counting the cells at the bottom of the inserts. (H) WT and Bim-KO Panc-1 cells were treated with 1 μM I-BET762, 5 μM GEM, and their combination for 72 h. Cell viability was determined by the CCK-8 assay. (I) WT and Bim-KO Panc-1 cells were treated with 1 μM I-BET762, 5 μM GEM, and their combination for 24 h. Apoptosis was detected by flow cytometry. The results of (B,C) and (G) are expressed as the means ± SD of 3 independent experiments. **P < 0.01.