E2F4 is a transcription factor with a prominent role in cell homeostasis. We have shown that a mutant form of mouse E2F4 (E2F4DN), carrying T249A/T251A mutations, is a multifactorial therapeutic agent against Alzheimer’s disease (AD). Our current hypothesis for the mechanism of action of E2F4DN is that the phosphorylation of the Thr249/Thr251 motif by the stress kinase p38MAPK prevents the homeostatic function of E2F4, while E2F4DN restores it. To test this hypothesis, we have focused on N2a neuroblastoma-derived neurons treated with camptothecin (CPT) to induce DNA damage, as a mechanism of cell stress followed by cell death. Here we show that p38MAPK activity becomes increased 6 h after CPT treatment. This correlates with the phosphorylation of E2F4 in Thr249, which is prevented by the p38MAPK-specific inhibitor SB203580. The significance of E2F4 phosphorylation in apoptosis was tested by overexpressing wild-type E2F4 (E2F4WT), a phosphomimetic form of E2F4 carrying Thr249Glu/Thr251Glu mutations (E2F4CA) or E2F4DN, followed by quantification of caspase3 cleavage. E2F1, which was used as a positive control, induced a statistically-significant increase of procaspase-3 cleavage 32 h after CPT treatment. This was mimicked by either E2F4WT or E2F4CA, while E2F4DN expression inhibited this effect. Pretreatment with SB203580 was able to prevent cell death in E2F4WT but not in E2F4CA, confirming the requirement for cell death of E2F4 phosphorylation. We conclude that the phosphorylation of E2F4 facilitates the death of N2a neurons carrying DNA damage, supporting the hypothesis that neuronal expression of E2F4DN maintains the homeostatic function of E2F4 under stress conditions.