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

Figure 3. H486R mutant does not inhibit TNFα- induced NF-κB activation.

Figure 3

HeLa cells were transfected with optineurin or its H486R mutant (25, 50 or 100 ng) along with NF-κB reporter plasmid and, after 22 h, treated with TNFα for 4 h. Luciferase activities relative to untreated control are shown (n = 4). Western blot shows expression of optineurin and its H486R mutant using HA tag antibody. HeLa cells were infected with adenoviruses for expressing HA tagged optineurin (Optn-AdV) or H486R (H486R-AdV) mutant or control virus (AdC). After 36 hours of infection, the cells were treated with TNFα for 5 min or left untreated. Cell lysates were then prepared for western blotting with antibodies for IκBα, HA tag and actin (loading control). GFP expression was used to monitor infection by adenoviruses. RGC-5 cells were transfected with optineurin or its H486R mutant (100 ng) along with NF-κB reporter plasmid and, after 22 h of transfection, treated with TNFα for 3 h. Luciferase activities relative to untreated control are shown (n = 4). Western blot shows expression of optineurin and its H486R mutant using HA tag antibody.