Ab13259

Western blotting was performed as described in our previous studies 55 (link). In brief, mice were killed by overdose of ketamine and xylazine, and dentate gyrus was isolated as described as described by Hagihara et al 56 . Proteins were extracted using RIPA buffer (P0013B, Beyotime). Protein concentration was determined by BCA method. An equal amount of total proteins for each sample was loaded, separated by 10% SDS-polyacrylamide gel electrophoresis, and transferred onto nitrocellulose membranes. The membranes were blocked with 5% defatted milk for 1 hour at room temperature, and subsequently incubated with primary and IRDye® 800CW-conjuncted secondary antibodies (1:2000, LI-COR Biosciences), in turn, overnight at 4 °C: GSK-3β (Phospho-Ser9) (1:1000, 9323S, Cell signaling), GSK-3β (Phospho-Tyr216) (1:1000,05-413, Millipore), GSK-3β (1:1000, 40988, Signalway Antibody), CDK5 (1:500, 33178, Signalway Antibody), DCX (1:1000, ab18723, Abcam), Synapsin1 (1:1000, ab64581, Abcam), synaptotagamin (1:1000, ab13259, Abcam), synaptophysin (1:1000, ab32127, Abcam), PSD95 (1:1000, 2507, Cell signaling), β-actin (1:1000, 21800, Signalway Antibody). The bands were visualized after washing with TBST using an Odyssey Infrared Imaging System (LI-COR biosciences, Lincoln, NE, USA). Bands were analyzed by Image J software (Fiji).