p.) 1 hr later. To assure see more similarity of SE intensity, we quantified behavioral and EEG seizures after infusion of KA and for 1 hr intervals after treatment with diazepam and lorazepam in both vehicle- and 1NMPP1-treated TrkBF616A mice ( Figures S3 and S4). The EEG recording electrode was placed in the left hippocampus so as not to confound histological

analyses of the hippocampus ipsilateral to the infused (right) amygdala; the extensive commissural connections between the hippocampi notwithstanding, it is possible that electrographic seizure activity localized to the right hippocampus occurred and escaped detection. Unless specified otherwise, after SE, animals underwent continuous video-EEG monitoring 24 hr/day, 7 days/week during weeks 1–2 and weeks 5–6 post-SE. Spontaneous recurrent seizures (SRSs) were identified by review of video-EEG files by two independent trained readers blinded to both genotype and treatment of mice. Behavioral seizures were classified according

to a modification of the Racine scale for mice (Borges et al., 2003). All EEG SRSs were confirmed by corresponding behavioral seizures documented by time-locked video review. Quantitative analysis of EEG energy content was performed as Birinapant cost described in Lehmkuhle et al. (2009) (Figures S3 and S4). In experiments examining effects of 1NMPP1 treatment on SE-induced spontaneous recurrent seizures, the first dose of 1NMPP1 (16.6 μg/g, i.p.) was injected immediately after giving diazepam and a second dose of 1NMPP1 (16.6 ng/g) immediately after administration

of lorazepam (Figure S1B). A third dose of 1NMPP1 (16.6 μg/g, i.p.) was injected approximately 12 hr post-SE, after which 1NMPP1 was administered daily (16.6 μg/g, i.p.) and also included in drinking water (25 μM) for the ensuing 2 weeks, at which point it was tapered and discontinued. WT mice and TrkBF616A mice injected under the same regimen with vehicle (i.p. and in drinking either water) served as controls. Animals were euthanized and decapitated. Crude membranes were prepared from hippocampi and subjected to SDS-PAGE. After transfer, western blotting was conducted as described in the Supplemental Experimental Procedures. After EEG and behavioral monitoring, KA-infused mice were examined for spontaneous activity in the open field and anxiety-like behavior in the light/dark box at 8 weeks post-SE as described in the Supplemental Experimental Procedures. PBS-infused (amygdala) WT or TrkBF616A mice treated with vehicle or 1NMPP1 were tested at 8 weeks postinfusion and served as controls. At 10 weeks post-SE, mice were anesthetized and perfused with heparinized PBS followed by 4% paraformaldehyde and brains prepared for immunofluorescent study of neurons and astrocytes as described by Mouri et al. (2008).