FIG 8.

Role of PI3K in regulating antiapoptotic and proapoptotic proteins in E. faecalis-infected cells. (A, B, and C) Lysates from infected RAW264.7 cells were prepared for Western blot analysis to detect the relative levels of Bax and Bcl-2 proteins using anti-Bax and anti-Bcl-2 antibodies, respectively. (D, E, and F) Untreated RAW264.7 cells or cells pretreated for 30 min with the PI3K inhibitor LY294002 (20 μM) were infected with E. faecalis for 1 h, and then the cells were washed and replenished with fresh medium containing vancomycin plus gentamicin and LY294002 (20 μM) for 12 h. The cells were harvested for Western blot analysis using Bax- and Bcl-2-specific antibodies. The relative densities of Bax and Bcl-2 protein bands on Western blots were compared to that of actin in each group, and fold activations relative to untreated samples were determined. Each data point in the graph represents the mean and SD of three independent experiments. *, P < 0.05 compared with control; ns, not significant compared with control.