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. Author manuscript; available in PMC: 2009 Dec 26.
Published in final edited form as: Cell. 2008 Dec 26;135(7):1311–1323. doi: 10.1016/j.cell.2008.10.044

Figure 3. CYLD knockdown inhibits necroptosis.

Figure 3

(A) L929 cells were transfected with 3 different siRNAs against cyld (cyld9 & cyld11), and rip1 (50nM) for 48hr, treated with 20μM zVAD.fmk for additional 18hr and the viability was measured as in Figure 1. Knockdown efficiency of cyld was confirmed by western blot using anti-CYLD, anti-RIP1 or anti-Tubulin antibodies. (B) Human FADD-deficient Jurkat cells were electroporated with siRNAs against human cyld (cyld6 & cyld7). Forty-eight hrs later, the cells were treated with 10ng/ml TNFα for additional 16hr and the viability was measured as in Figure 1. The cell lysates were analyzed by western blotting as in (A). (C) Inhibition of LC3-II induction in cyld-knockdown cells. L929 cells stably expressing shRNA for cyld(sh-cyld) or vector alone(vector) were treated with 20uM zVAD.fmk for indicated periods and the cell lysates were analyzed by western blotting using anti-LC3-II, anti-CYLD or anti-Tubulin antibodies.