Fig. 1

Design of shRNA targeting Sirt1 for AAV packaging and hippocampal Sirt1 knockdown by stereotactic injection. A Validation of knockdown efficiency of Sirt1-shRNAs in GL261 cells by RT-qPCR. Abbreviations: GL261-WT (wide type GL261 cells), GL261-shSirt1A (GL261 cells infected by the first shRNA sequence targeting Sirt1), GL261-shSirt1B (GL261 cells infected by the second shRNA sequence targeting Sirt1). n = 3. ^***p < 0.001 represents comparison between the GL261-WT and GL261-shSirt1A. ^#p < 0.05 represents comparison between the GL261-WT and GL261-shSirt1B. B Western blotting of Sirt1 levels after Sirt1 knockdown in G1261 cells. (C) Quantitative density values in B. D Illustration of plasmid construction for AAV packaging. Abbreviations: inverted terminal repeats (ITR), cytomegalovirus promoter (CMV), Sirt1 shRNA (shSirt1), and enhanced green fluorescent protein (EGFP) reporter gene. E Work flow of in vivo experiments performed on the control group, the sham group and the shSirt1 group. Abbreviations: Open Field test (OFT), Morris Water Maze (MWM), magnetic resonance imaging (MRI), and western blotting (WB). F Graphical illustration of stereotactic injection of AAV into bilateral CA1 regions of hippocampus. G Representative fluorescence image of frozen brain section