Experimental schematic and transduction of hippocampal neurons with AAV1-Rheb(S16H) in SD rats. (A) Experimental schematic for investigating the effects of AAV1-Rheb(S16H) administration in the rat hippocampus in vivo. (B) Double immunofluorescence staining to visualize co-expression patterns of FLAG and NeuN, or FLAG and GFAP in the Rheb(S16H)-transduced hippocampus. Scale bar, 50μm. The graph represents the ratio of FLAG-colocalized cells between neurons and non-neuronal cells in the AAV1-Rheb(S16H)-injectied hippocampus (n=3 per group). (C) Immunohistochemical staining of BDNF expression following to Rheb(S16H) transduction of hippocampal neurons in vivo. Lower images are magnified photomicrographs of the insets. Scale bars: upper images, 100 μm; lower images (insets), 20 μm. (D) Western blot analysis for BDNF expression in the controls, AAV1-GFP- and AAV1-Rheb(S16H)-injected hippocampus of rat brains. Differences among groups were evaluated by one-way analysis of variance and Tukey’s post-hoc test (*p<0.05 vs. CON; n=3 per group).