Figure 3.
ROS are generated during caspase-independent death. A, B, E18 cortical neuron cultures were treated with no additives (ctrl) or with the combination of camptothecin and BAF (cam+BAF) for 20 hr. Generation of ROS was assessed by incubation with the fluorescent dye DHE (red). Neurons were counterstained with the green fluorescent viability indicator calcein AM (A). Alternatively, the green fluorescing probe DCF in combination with the death indicator ethidium homodimer (eth. h., red) was used (B). The percentage of DHE-positive neurons relative to live (calcein AM-positive) neurons was counted over time. In parallel, neuronal survival was assessed by nuclear counts. Numbers are expressed as mean ± SEM (n = 3).