Figure 2

Increased NF-κB luciferase activity in neuronal cells with knockdown of RNF11. (A) Stable SH-SY5Y cell lines were generated using lentiviruses containing a control scramble small hairpin RNA (shRNA) sequence (shRNA-Scramble cells) or shRNA targeted against RNF11 (shRNA-RNF11 cells). Quantitative RT-PCR was used to measure relative RNF11 mRNA levels with glucuronidase β as an internal control. Values are expressed as mean comparative cycle threshold ± SEM of triplicate experiments. (B) Stable SH-SY5Y cell lines, as well as untransduced cells, were transiently cotransfected with luciferase and Renilla plasmids and stimulated with 10 ng/ml TNF-α or PBS for 6 hours. Values represent the fold change in NF-κB-dependent activity as measured by the ratio of luciferase to Renilla luminescence in stimulated over unstimulated samples ± SEM in triplicate experiments. ^* P < 0.05; ^*** P < 0.001.