A) Cartoon depicting the pro-survival and pro-death arms of the TNF signaling pathway.
B) FLAG IP of the TNFR-SC in hTERTs following a 15 min stimulation with 1 μg/mL FLAG-TNF.
C) hTERTs stimulated with TNF over 24 hrs (n = 3).
D) hTERTs stimulated with TNF assessed for loss of pCYLD. Phosphorylation at this site decreases deubiquitinase activity.
E) hTERTs pretreated for 30 min with either birinapant and zVAD, or birinapant, zVAD, and Nec-1 were exposed to TNF for 24 hrs. Relative viability was measured by resazurin reduction (n = 3).
F – G) hTERTs complemented with DOX-inducible TBK1 were pretreated for 30 min as indicated and then stimulated with TNF (5 ng/mL) for 24 hrs. Relative viability was measured by resazurin reduction (F n = 3, G n = 6).