synaptotagmin 1, Santa Cruz Biotechnology, catalog #sc-136480; PSD95, Synaptic systems, catalog #124011; syntaxin, I378, custom made, gift of T. Südhof) overnight at 4°C. The following day, the blots were washed three times with tris-buffered saline with 0.1% Tween 20 (TBST) before incubating with horseradish peroxidase–conjugated secondary antibody (1:5000) at room temperature for 1 hour. After four washes with TBST, the blots were incubated with clarity Western ECL substrate (Bio-Rad, catalog #1705060) for 1 min before developing the membrane on autoradiographic films (Lab Scientific, catalog #XAR ALF 1318).
