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. 2022 Jan 12;13(1):52. doi: 10.1038/s41419-021-04483-0

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© The Author(s) 2022

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 — Flag TNF-induced complex-I immunoprecipitation. MC-38 cells (A) or HT-29 cells (B) were stimulated with Flag-TNF (1 µg/mL) for the indicated time points, followed by Flag immunoprecipitation and immunoblot analysis. HT-29 cells were pre-treated with 1 nM LCL161 for 1 h before Flag-TNF stimulation cell lysates of MC-38 cells (C) or HT-29 cells (D) stimulated with or without TNF were immunoprecipitated with antibody against RIPK1 or antibody against SMYD2, followed by immunoblot analysis. HT-29 cells were pre-treated with 1 nM LCL161 for 1 h before TNF stimulation. E Immunoblot of phosphorylated or total RIPK1 in lysates of WT and SMYD2 deficient HT-29 cells stimulated with TS (Upper panel) or TSZ (Lower panel) at different time points. β-Actin served as a loading control. F A schematic diagram depicting SMYD2 mediated cell death pathways in CRC cells.