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

Figure 6. Optineurin is required for deubiquitination of RIP by CYLD.

Figure 6

HeLa cells were infected with adenoviruses expressing shRNA against optineurin or with control adenoviruses. After 48 hrs of infection, cells were either left untransfected or transfected with HA- CYLD, treated with TNFα for 10 min after 30 h of transfection and subjected to immunoprecipitation with RIP or control antibodies. Immunoprecipitates were then subjected to western blotting with RIP antibody. The blot was then reprobed with Lys63-linked polyubiquitin (K63-Ub) antibody. HeLa cells were infected with adenoviruses expressing wild-type optineurin or H486R mutant. After 4 h of infection, cells were either left untransfected or transfected with HA- CYLD, treated with TNFα for 10 min after 24 h of transfection and subjected to immunoprecipitation with RIP or control antibodies. Immunoprecipitates were then subjected to western blotting with RIP antibody. The blot was then reprobed with Lys63-linked polyubiquitin (K63-Ub) antibody.