E coli that express HlyA invoke Bcl-xL protein degradation in platelets. (A) Platelets were incubated with wild-type UTI89 or UTI89 ΔhlyA bacteria for 4 or 8 hours and Bcl-xL and actin protein were assessed by Western blot analysis. (B) Platelets were incubated with wild-type UTI89 or UTI89 ΔhlyA bacteria for 4 or 8 hours and intracellular Bcl-xL protein was quantified by ELISA. The bars in the graph are the mean ± SEM (n = 3) and the single asterisk indicates a significant decrease (P < .01) in Bcl-xL protein levels compared with all other conditions. (C) Platelets were incubated with WAM582 or WAM783 for 2, 4, or 8 hours, and Bcl-xL protein was assessed by ELISA. The bars in the graph are the mean ± SEM (n = 3), and the asterisk indicates a significant decrease (P < .01) in Bcl-xL protein levels in WAM582 treated platelets compared with all other conditions. (D) Platelets were incubated with wild-type UTI89, UTI89 ΔhlyA, WAM783, WAM582, and S aureus isolated from the bloodstream of a patient diagnosed with sepsis (SA-SP), α-toxin (500 ng/mL), or thrombin (1 U/mL). After 8 hours, protein for Bcl-xL and actin were assessed by Western blot analysis. The Western blots in this figure are representative of 2 independent experiments for each group.
