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. 2016 Sep 29;7(9):e2381. doi: 10.1038/cddis.2016.294

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Copyright © 2016 The Author(s)

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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The adaptor protein TRADD is indispensible for caspase 8 activation and necrosome formation in the setting of TAK1 inhibition. (a) Western blots for the indicated proteins from TNFR1-/-, and TNFR2-/- MEFs treated with vehicle control or 5z-7 plus TNFα for 4 h, in the presence or absence of Nec-1 or zVAD. (b) Caspase 8 activity in TNFR1-/-, and TNFR2-/-, MEFs treated with vehicle control, TNFα, or 5z-7 plus TNFα for 2 h. †P<0.01 versus Control or TNF only. (c) Western blots for the indicated proteins from TRADD+/+ and TRADD-/-, MEFs treated with vehicle control or 5z-7 plus TNFα for 4 h, in the presence or absence of Nec-1 or zVAD. (d) Western blots for the indicated proteins following IP with an anti-RIP3 antibody from TRADD+/+ and TRADD-/-, MEFs treated with 5z-7 plus TNFα for the indicated periods of time. (e) Caspase 8 activity in TRADD+/+ and TRADD-/-, MEFs treated with vehicle control or 5z-7 plus TNFα for 2 h, with or without Nec-1 or zVAD. †P<0.01 versus Con; ^#P<0.05 versus corresponding TRADD+/+. (f) Cell death assessed by PI staining of TRADD+/+ and TRADD-/-, MEFs treated as in e for 4 h. ^†P<0.01 versus Con; ^#P<0.05 versus corresponding TRADD+/+ ^§P<0.05 versus TRADD+/+ 5z-7 plus TNF. * indicates cleaved proteins