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. Author manuscript; available in PMC: 2016 Jun 15.
Published in final edited form as: Cell Rep. 2016 Jun 2;15(11):2449–2461. doi: 10.1016/j.celrep.2016.05.032

FIGURE 4. Elimination of CYLD protects macrophages from TLR4-induced auto-necroptosis.

FIGURE 4

(A) BMDMs from Cyldflox/flox (CYLD WT) and LysM-Cre x Cyldflox/flox mice (CYLD conditional knockout, or CKO) were pre-treated with DMSO or 25 μM zVAD-fmk and stimulated with LPS for 16 hours. (B) BMDMs from CYLD WT or CYLD CKO were pre-treated with 25 μM zVAD-fmk prior to infection with E. coli or C. rodentium for 16 hours at the indicated MOI. (A–B) Cell death was analyzed as in Figure 3A. * p<0.05, ** p<0.01, *** p<0.001. (C) Cyld+/+ or Cyld−/− BMDMs pre-treated with 25 μM zVAD-fmk were stimulated with 1 ng/ml of LPS for the indicated time. Triton-soluble lysates were subject to non-reducing SDS-PAGE, and blotted with anti-MLKL and β-actin. The experiment shown is representative of three independent experiments.