Figure 12.

DNA damage-induced death of CGNs is associated with caspase-dependent p25 production and sensitization by DN-NES. A, Western blot analysis of total cellular protein from CGNs treated 2 d after plating with camptothecin (campto) alone or camptothecin plus BAF (100 μm) for the indicated times. Quantification of relative p35 and p25 levels is shown on the right. Density values for each band were normalized to actin and then to the control time point. B, CGNs treated with camptothecin alone or camptothecin plus BAF (100 μm) were assayed for survival at the indicated times according to the protocol outlined in Materials and Methods. C, CGNs were infected at the time of plating with a multiplicity of infection of 20. At 48 h after plating/infection, neurons were treated with 10 μm camptothecin for the indicated times. Survival was assessed after 24 h of camptothecin treatment. GFP-positive neurons were classified as either dead or alive according to the appearance of Hoescht. The ratio of live to total cells counted was used as a measure of survival. Error bars represent the mean ± SEM for data from three independent experiments each done in triplicate. *p < 0.05 by ANOVA. A minimum of 100 cells were counted per well. Infected cultures not treated with camptothecin showed low toxicity (>85% survival for all constructs).