DAPK1-deficient mice display a significant delay in the development of kindling. Mice (C57BL/6, male, P56-P80, n = 12/group) were treated with a subconvulsive dose of PTZ (35 mg/kg) every other day (nine injections for 17 days). The brain tissues from a number of the animals (n = 4) were harvested after the fourth PTZ injection, and the remaining animals continued to undergo behavioral analysis. A The mean seizure grades of PTZ-kindled WT and DAPK1 KO mice; DAPK1 KO mice exhibited significantly prolonged latency to the first kindled seizure scored ≥ 4 (**P < 0.01, one-way ANOVA with Tukey's multiple comparisons test). B Latency to the first kindled seizure was significantly prolonged in the DAPK1-deficient mouse group (***P < 0.001, two-tailed Student's t test). C-E Enhanced interaction between NR2B and DAPK1 in the hippocampus after PTZ exposure (***P < 0.001 vs control, two-tailed Student's t tests). F-H Brain lysates were analyzed via immunoblotting with anti-pSer1303-NR2B, anti-NR2B, anti-pSer308-DAPK1, anti-DAPK1, or anti-β-actin antibodies. Statistical significance was determined by one-way ANOVA with Tukey's multiple comparisons tests (**P < 0.01 and ***P < 0.001). n.s., no significance. All data shown represent the means ± standard errors of three independent experiments.