Animal procedures
Preparation of cortical neurons
Cell culture and transfections
Biotin-switch technique
S-nitrosothiol resin-assisted capture
Coimmunoprecipitation
Western blotting
Dendritogenesis assay
Plasmids
GO analysis
Mass spectrometry
SNO-site identification
Protein identification from neuronal extracts
NanoLC-MS/MS analysis of neuronal extracts
Analysis of neuronal extract data
Hit criteria for SNO-Ps
Motif-X analysis
Statistical analysis
Standard silver staining protocol was followed. Briefly, the polyacrylamide gel was soaked in 50% methanol two times for 15 min and then in 5% methanol for 10 min before rinsing in water. Gel was then soaked in 10 μM DTT for 20 min and then in 0.1% silver nitrate for 20 min. After three washes with water, gel was washed two times in developer solution (500 ml of ~285 mM sodium carbonate solution plus 250 μl of 37% formaldehyde) and soaked until bands became visible. The reaction was stopped with addition of citric acid, and gel was washed in water before imaging.