Coronal brain sections (40-µm thick) were prepared from injected mice and processed for immunostaining as described (Ge et al., 2006). Images were acquired on a META multiphoton confocal system (Zeiss LSM 510) using a multi-track configuration. Analysis of neuronal development and pAKT/pS6 were performed as previously described (Duan et al., 2007; Kim et al., 2009). All experiments were carried out in a blind fashion to experimental conditions. Statistic significance was determined by ANOVA or student t-test as indicated.
