FIG 6.

Blockade of PI3K activation reverses E. faecalis-induced antiapoptotic activity. (A) RAW264.7 cells were infected with E. faecalis E99 as described in Materials and Methods and analyzed for pAkt levels at 6, 12, and 24 h by Western blotting. (B) RAW264.7 cells were pretreated with LY294002 (20 μM) for 30 min and then infected with enterococcus (MOI = 10). After 1 h of incubation, the cells were washed and replenished with fresh medium containing vancomycin plus gentamicin and LY294002 (20 μM) for 12 h and then treated with STS to induce apoptosis. TUNEL staining was used to measure cell apoptosis. (C) Cell lysates after different treatments were harvested for Western blotting. Cleaved caspase 3 levels were normalized by actin densitometry, 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; ns, not significant.