Pseudomonas aeruginosa ExoT Induces Atypical Anoikis Apoptosis in Target Host Cells by Transforming Crk Adaptor Protein into a Cytotoxin
Fig 5
Crk mediates ExoT-induced apoptosis.
Crk-/- cells were transiently transfected with pCrkI, or pGFP on 3 consecutive days to obtain more transfected cells. 24 hr after the final transfection, cells were infected with either ∆U, ∆U/T(G-A+), or pscJ at MOI~10. Cytotoxicity of transfected host cells (green) was assessed by fluorescent time-lapse microscopy, using PI uptake (red cells are dead). Cytotoxicity is expressed as a percentage of the total number of transfected cells. Video images were captured every 15 min and selected movie frames at indicated time points are shown in (A). (For clarity, phase panels were excluded from the merged images). The corresponding tabulated data are shown in (B). (C) The corresponding time to death, defined as the time of infection to the time of PI uptake (red) is expressed as the mean ± SEM. The data in B and C comprise 3 independent experiments. Statistical analysis was performed using one-way ANOVA.