Local Oxidative Stress Expansion through Endothelial Cells – A Key Role for Gap Junction Intercellular Communication
Figure 6
Apoptotic cell death in bystander cells following COI.
A – bEnd.3 monolayers were linearly scratched by a surgical scalpel (arrow), subjected to COI (white rectangle, left of scratch upper panel) and placed back in the incubator for 3h. Then, CaspACE FITC-VAD-FMK, PI and DAPI were added to the cells. Upper and lower panels are snapshots of the same field of view. Middle panel is a higher magnification snapshot of the yellow rectangle in upper panel. Cell death (PI+) is preceded by CaspACE staining at 100–200µm beyond the COI, left of the scratch only. B – bEnd.3 monolayers were placed in the incubator after COI for 3h, and then subjected to Annexin-V-FITC (for phosphatidylserine depolarization) and PI. Phosphatidylserine depolarization ((Annexin-V)+ cells (green)) clearly precedes cell death and permeabilization (PI+). Panels show different magnifications. Images are a representative of n = 3 experiments.