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. Author manuscript; available in PMC: 2012 Jun 1.
Published in final edited form as: Nat Cell Biol. 2011 Oct 30;13(12):1437–1442. doi: 10.1038/ncb2362

Figure 4. CYLD-D215A triggers programmed necrosis in the absence of caspase inhibitors.

Figure 4

(a) CYLD-WT and CYLD-D215A MEFs from Figure 3d undergoing necrosis 24 hours after stimulation with TNF in the presence of IETD-fmk or Necrostatin-1 (NEC1) was quantified by Annexin V staining. (b) Bar chart displays the mean percentage of CYLD-WT and CYLD-D215A cells cultured in triplicate undergoing necrosis in the presence of vehicle or zVAD-fmk after TNF stimulation for 24 hours. Error bars display standard deviation and * denotes p = 0.0000446 (n=3). (c) Cyld−/− MEFs reconstituted with CYLD-D215A were depleted of RIPK1 and RIPK3 for 48h prior to TNF stimulation for 24 hours to induce necrosis. Immunoblot confirms knockdown of RIPK1 and RIPK3.