Figure 3

LAIR-1 engagement inhibits CpG-mediated nuclear translocation of IRFs and the activation of NF-κB. Human monocytes were stimulated with CpG, C1q or anti-LAIR-1 antibody for 3 h. Nuclear fractions were analyzed by Western blot for IRF5, IRF3 and lamin for a marker for the nuclear fraction. C1q and anti-LAIR-1 antibody inhibit CpG-mediated nuclear translocation of IRF5 and IRF3 (A). Data are representative of three independent experiments. (B) Stimulated cells were stained for IRF3 and propodium iodine (PI) as a nuclear marker. The percentage of IRF3 nuclear translocation was assessed in more than 200 cells in three independent experiments, and representative images are shown. All images were viewed at 120× magnification by using a FluoView 300 confocal microscope (Olympus). (C) Purified monocytes were stimulated with CpG or C1q together with CpG for the indicated times, and cell lysates were assayed for phosphorylated-IkBα by Western blotting. Phosphorylated IkBα was normalized to GAPDH. Experiments were repeated three times, and representative data are shown.