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. 2018 Sep 17;9(10):935. doi: 10.1038/s41419-018-0960-8

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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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Fig. 4 — a The Bim expression in Bim-KO cells were analyzed by western blotting. b WT and Bim-KO HepG2 cells were treated with idelalisib at indicated concentration for 24 h. Apoptosis was analyzed by a nuclear fragmentation assay. The results were expressed as the means ± SD of three independent experiments. **P < 0.01; *P < 0.05 (one-way ANOVA with Tukey’s post hoc test). c WT and Bim-KO HepG2 cells were treated with 5 μmol/L idelalisib for 24 h. Apoptosis was analyzed by annexin V/PI staining followed by flow cytometry. The results were expressed as the means ± SD of three independent experiments. **P < 0.01 (one-way ANOVA with Tukey’s post hoc test). d WT and Bim-KO HepG2 cells were treated with 5 μmol/L idelalisib for 24 h. Cleaved caspase 3 and 9 expression was analyzed by western blotting and normalized to β-actin. Data represent the mean ± SD of three independent experiments. *P < 0.05 (Student’s t-test). (e) The cytoplasm and mitochondria were fractionated from WT and Bim-KO HepG2 treated with 5 μmol/L idelalisib for 24 h. The distribution of cytochrome c was analyzed by western blotting and normalized to β-actin. Data represent the mean ± SD of three independent experiments. *P < 0.05 (Student’s t test). β-actin and cytochrome oxidase subunit IV (Cox IV) were analyzed as the control for loading and fractionation. f WT and Bim-KO HepG2 cells were treated with 5 μmol/L idelalisib for 24 h. Colony formation assay was done by seeding an equal number of treated cells in 12-well plates, and then staining attached cells with crystal violet 14 days later. Left, representative pictures of colonies; Right, quantification of colony numbers. The results were expressed as the means ± SD of three independent experiments. **P < 0.01 (Student’s t-test)