FIGURE 2. TLR4-TRIF signaling and not TNF mediates CASPASE-8 dependent cleavage of CYLD.
(A) Wild type and Tnf−/− BMDMs were treated with 100 ng/ml of LPS for 4 hours. (B–C) Wild type, Myd88−/−, and Trif−/− BMDMs were treated with 10 ng/ml of LPS for the indicated times. (D) Wild type BMDMs were pre-treated with 30 μM Necrostatin-1 prior to stimulation with LPS for 4 hours. (E) Immortalized BMDMs generated from RosaCreERT2Ripk1fl/fl mice were treated with ethanol or with tamoxifen (4-OHT) to delete RIPK1. Cells were then treated with 100 ng/ml of LPS for the indicated times. Lysates were blotted as indicated. All experiments shown are representative of at least three independent experiments with similar results.