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. 2011 Mar 7;6(3):e17477. doi: 10.1371/journal.pone.0017477

Figure 5. Optineurin is required for CYLD mediated inhibition of NF-κB activation.

Figure 5

HeLa cells were infected with adenoviruses expressing shRNA against optineurin or with control adenoviruses. After 48 hrs of infection cells were transfected with NF-κB-Luc with or without CYLD (100 ng). The cells were treated with TNFα for 4 h after 22 h of transfection. Luciferase activities relative to untreated control are shown (n = 4). Western blot shows the expression of HA-CYLD in control and optineurin knockdown cells. RIP ubiquitination is enhanced in optineurin knockdown cells. HeLa cells were infected with Ad-shOPTN or with control (Cont-AdV) adenoviruses. After 72 h of infection cell lysates were subjected to immunoprecipitation with RIP antibody or control antibody. The immunoprecipitates were analyzed by western blotting. ns, non-specific. * indicates ubiquitinated RIP.