Beclin 1 is required for neuronal survival in vitro. a Representative western blot of lysates from primary forebrain neurons isolated from wild type E16.5 embryos. Neurons were infected at 1 week (wk) in culture with a lentivirus encoding a control scrambled or beclin 1-targeted shRNA (ctrl or bec shRNA, respectively) as well as copGFP. b Quantification of beclin 1 levels relative to the loading control neuron specific enolase (NSE). Data are combined results of three independent experiments. Data were analyzed by unpaired Student’s t-test. c Timeline of survival experiment. d Representative images of neurons at the 3weeks post-infection (P.I.) time point. Infected cells are GFP positive. Neurons were stained for MAP2 (red) and Hoechst’s dye (blue). e Quantification of MAP2+ cells at 2 and 3 weeks P.I. The number of surviving MAP2+ cells is expressed relative to the number of cells in the ctrl shRNA wells. Data are combined results from 3 independent experiments (n = 36 fields/group). Data were analyzed by two-way ANOVA with Sidak’s post-test. ****p ≤ 0.0001., **p ≤ 0.01, ****p ≤ 0.0001. Bar graphs are mean ± SEM.