FIGURE 10.

Attenuation of Eps8, Src, or FAK decreases bacterium killing activity of macrophages. A, equal numbers (5 × 10⁵) of RAW cells and their derived Ctrl cells, Eps8-attenuated cells (siEps8-346-1), Src-attenuated cells (siRNA-1), Src-attenuated cells expressing avian c-Src (siRNA-1/Src6), or FAK-attenuated cells (fak siRNA-1) were pretreated with PBS (−) or LPS (100 ng/ml) for 24 h. The assessment of phagocytosis of E. coli and the following bacterium killing in these cells is described under “Experimental Procedures.” B, PEMs (1 × 10⁶) prepared from C57BL/6 mice were infected with lentivirus (MOI = 3) encoding luciferase siRNA (Ctrl) or eps8 siRNA (siEps8-503). After 2 days, PEMs were stimulated without (−) or with LPS (100 ng/ml) for 24 h. The lysates (100 μg) were resolved by SDS-PAGE and probed with antibodies as indicated. The arrow indicates the position of active Src. Bacterium killing activity of Ctrl and siEps8-503 cells was performed exactly as described above. The results in the A and B are means ± S.D. (error bars). *, p < 0.05; **, p < 0.01, as compared with the corresponding control. Experiments were repeated three times in triplicate, and representative results are shown.