Neurons and cells were lysed in RIPA buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton-X 100, plus a cocktail protease inhibitors (Roche)), and lysates were analyzed by SDS-PAGE in the presence of DTT (0.1 M). Immunoblotting and quantitative analysis were performed by a dual-channel infrared imaging system with fluorescence-labeled secondary antibodies (800CW and 680LT), an Odyssey Infrared Imager CLX and software Image Studio 5.2.5 (LI-COR Biosciences). Signals were normalized for Tuj1 probed on the same blots as loading controls. Antibodies used were listed in Key Resources Table.
