Cells were fixed at room temperature for 20-30 min with 4% formaldehyde/4% sucrose in PBS, permeabilized with 0.25% Triton-X in PBS, and blocked with 2% BSA/4% goat serum in PBS. The following primary antibodies were used: mouse anti-microtubule-associated protein 2 (MAP2) monoclonal (1:1000; Sigma; M4403), rabbit anti-MAP2 polyclonal (1:1000; Chemicon; AB5622); mouse anti-bassoon monoclonal (1:1000; Stressgen Bioreagents; VAM-PS003), anti-GAD67 (1:1000; Millipore; MAB5406), chicken anti-GFP polyclonal (1:500; Aves Lab; GFP-1020), and mouse anti-phospho-CREB (Ser133) monoclonal (1:1000; Millipore; 05-807). For secondary antibodies, we used anti-mouse, anti-rabbit, or anti-chicken conjugated Alexa Fluor 488, 568, or 647 (1:500; Invitrogen).
