NO inhibits ribosome complex formation through phosphorylation of eIF-2α. (A) RAW264.7 cells were stimulated with LPS + IFN-γ for 14 hr. Cells were isolated and phosphorylation of eIF-2α was analyzed after VIEF and immunoblotting with a monoclonal antibody to eIF-2α. (B) Ribosome profiles were determined by isolation of the cytosolic fraction of RAW264.7 treated with LPS + IFN-γ for 24 hr, sucrose gradient centrifugation, and continuous measurement at an optical density of 280 nm. (C) Cells were treated with LPS + IFN-γ for 12 hr and then labeled with 35S-methionine for 2 hr in the presence or absence of 400 µM 8Br-cGMP. Data are expressed as mean ± SD; n ≥ 3.