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. 2018 Sep 20;50(9):125. doi: 10.1038/s12276-018-0152-8

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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. 3 — a TNF-induced necroptosis was completely blocked by HS-1371 treatment. HT-29 cells were pretreated with four tested inhibitors for 2 h and then treated with TSZ (6 h for immunoblotting, 24 h for cell death assay). Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay or phase-contrast microscopy. The results are presented as the means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. b Protection of necroptosis by HS-1371 treatment is similar with dabrafenib, which abolished downstream events. HT-29 cells were pretreated with various necroptosis inhibitors and HS-1371 before TSZ treatment. Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay (left panel). Cells were stained with phospho-MLKL antibody and analyzed by confocal fluorescence microscopy (middle panel), and cell death was further analyzed by FACS analysis after PI staining (right panel). c HS-1371 protects cells from TNF-induced necroptosis in a dose-dependent manner. HT-29 cells were pretreated with indicated concentrations of HS-1371 and then treated with TSZ (6 h for immunoblotting, 24 h for cell death assay). Cell lysates were analyzed by immunoblotting, and cell viability was analyzed by MTT assay or phase-contrast microscopy. The results are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001