Bmx does not signal upstream of NF-κB. Macrophages were infected for 2 hours with adenoviruses overexpressing wild-type Bmx or control adenovirus Ad0 in serum-free medium. For luciferase assays, cells were subjected to a second round of infections with NF-κB consensus luciferase adenovirus (AdNFκB-luc) before stimulation. Cells were stimulated with 10 ng/mL LPS for 18 hours for ELISA, 4 hours for luciferase reporter gene assays, and 0.5 and 1 hour for EMSA. (A) Uninfected, Ad0-infected, and AdBmx-infected macrophages were left untreated or preincubated with PSI (1 μM) and TNF and IL-6 production assessed by ELISA. Values (mean ± SEM) are presented as percent compared with LPS-treated uninfected controls (100%; *P< .05; **P< .01). (B) Relative luciferase units (RLU) for uninfected, Ad0-infected, and AdBmx-infected cells in the absence (left hand panel) or presence (right hand panel) of AdNFκB-luc after 4 hours of LPS stimulation are shown as mean (± SEM) for 4 separate donors normalized to LPS only controls. n.s, not significant. (C) Nuclear extracts were collected as described in “Electrophoretic mobility shift assays” and assayed for NF-κB DNA binding by EMSA. Statistical significance was assessed by Student ttest.